Matrix Biology 18 Ž1999.

211᎐223

Mini-review

Role of fibronectin-binding MSCRAMMs in bacterial

adherence and entry into mammalian cells

Danny Joha,1, Elisabeth R. Wanna , Bernd Kreikemeyer a,2 , Pietro Spezialeb,

Magnus Hook¨¨ a,U
a
Center for Extracellular Matrix Biology, Albert B. Alkek Institute of Biosciences and Technology, Texas A & M Uni¨ ersity System
Health Science Center, 2121 West Holcombe Boule¨ ard, Houston, TX 77030, USA
b
Department of Biochemistry, Uni¨ ersity of Pa¨ ia, 27100 Pa¨ ia, Italy

Received 11 February 1999; accepted 16 February 1999

Abstract

Most bacterial infections are initiated by the adherence of microorganisms to host tissues. This process involves the
interaction of specific bacterial surface structures, called adhesins, with host components. In this review, we discuss a group of
microbial adhesins known as Microbial Surface Components Recognizing Adhesive Matrix Molecules ŽMSCRAMMs. which
recognize and bind FN. The interaction of bacteria with FN is believed to contribute significantly to the virulence of a number
of microorganisms, including staphylococci and streptococci. Several FN-binding MSCRAMMs of staphylococci and strepto-
cocci exhibit a similar structural organization and mechanism of ligand recognition. The ligand-binding domain consists of
tandem repeats of a ; 45 amino acid long unit which bind to the 29-kDa N-terminal region of FN. The binding mechanism is
unusual in that the repeat units are unstructured and appear to undergo a conformational change upon ligand binding. Apart
from supporting bacterial adherence, FN is also involved in bacterial entry into non-phagocytic mammalian cells. A sandwich
model has been proposed in which FN forms a molecular bridge between MSCRAMMs on the bacterial surface and integrins
on the host cell. However, the precise mechanism of bacterial invasion and the roles of FN and integrins in this process have
yet to be fully elucidated. 䊚 1999 Elsevier Science B.V.rInternational Society of Matrix Biology. All rights reserved.

Keywords: Extracellular matrix; FN; MSCRAMM; Bacterial adherence; Bacterial invasion

Abbre¨ iations: ECM, extracellular matrix; ELISA, enzyme-linked immunosorbent assay; BHK, baby hamster kidney; FI, type I module in
FN; FN-N29, the N-terminal 29-kDa fragment of FN; GST, glutathione-S-transferase; MSCRAMM, microbial surface component recognizing
adhesive matrix molecules
U
Corresponding author. Tel.: q1-713-677-7551; fax: q1-713-677-7576.
E-mail address: mhook@ibt.tamu.edu ŽM. Hook ¨¨ .
1
Current address: Xenogen Corporation, 860 Atlantic Avenue, Alameda, CA 94501, USA
2
Current address: Department of Medical Microbiology and Immunology, University of Ulm, Robert-Koch-Strabe 8, D-89081, Ulm,
Germany

0945-053Xr99r$ - see front matter 䊚 1999 Elsevier Science B.V.rInternational Society of Matrix Biology. All rights reserved.
PII: S 0 9 4 5 - 0 5 3 X Ž 9 9 . 0 0 0 2 5 - 6
212 D. Joh et al. r Matrix Biology 18 (1999) 211᎐223
1. Introduction
The process of microbial infection involves a com-
plex series of events that can result in host tissue
malfunction or destruction. The adherence of the
microorganisms to host tissues represents a critical
first step in this process. Non-adherent bacteria can
be readily eliminated from the body by such cleansing
mechanisms as peristalsis and excretion. Bacteria that
adhere to host tissues may remain extracellular or
may be internalized into an intracellular compart-
ment. Cellular invasion enables the bacteria to escape
host defense mechanisms, persist in the host and
invade tissues surrounding the initial site of coloniza-
tion. As a result of its critical role in the infection
process, bacterial adherence to host tissues represents
a potential target for the development of new antimi-
crobial agents. Agents that can block adherence, such
as specific antibodies, may prove to be effective in
combating infectious disease. For example, active im-
munization of mice with a recombinant fragment of
the collagen-binding protein from Staphylococcus au-
reus, or passive immunization with antibodies against
this protein, protect against sepsis-induced death
ŽNilsson et al., 1998.. Also, mice actively immunized
with a recombinant decorin-binding protein from
Borrelia burgdorferi are immune to challenge in a
model of lyme borreliosis ŽHanson et al., 1998..
The adherence of bacteria involves bacterial sur-
face components, called adhesins, that recognize and
bind to host extracellular matrix ŽECM. and cell
surface molecules. Host ECM components that are
known to support bacterial adherence include fi-
bronectin ŽFN., collagen, fibrinogenrfibrin, elastin,
vitronectin, laminin, as well as decorin and heparan
sulfate-containing proteoglycans. The subfamily of
bacterial adhesins that bind to ECM molecules are
collectively known as Microbial Surface Compo-
nents Recognizing Adhesive Matrix Molecules
ŽMSCRAMMs. ŽPatti and Hook, ¨¨ 1994; Patti et al.,
1994.. Although a number of microbes have been
shown to bind host ECM components, the
MSCRAMMs responsible for these interactions have,
in most cases, not been identified or characterized.
However, a group of related FN-binding MSCRAMMs
from gram-positive bacteria has been analyzed in some
detail.
In this review, we will discuss the bacterial
MSCRAMMs that interact with FN. FN is a 440-kDa
mosaic glycoprotein which is present in soluble and
matrix forms in various body fluids and tissues. It is
composed of three types of modules, presented in two
similar disulfide-linked subunits, and contains discrete
binding sites for a variety of other extracellular
molecules, including fibrin, heparin and collagen, as
well as for a number of integrins, including ␣4b1,
␣ 5b1, ␣ IIbb3 and ␣ vb3.
2. FN-binding MSCRAMMs from staphylococci and
streptococci
Staphylococci and streptococci are clinically impor-
tant gram-positive bacteria that are capable of caus-
ing a wide variety of diseases in humans and animals.
The specific binding of FN to Staphylococcus aureus
was first reported by Kuusela Ž1978.. Subsequently,
two FN-binding proteins ŽFnbpA and B. and the
corresponding genes were isolated and characterized
¨
ŽFlock et al., 1987; Froman ¨ et al.,
et al., 1987; Signas
¨
1989; Jonsson et al., 1991.. Many staphylococci and
streptococci have since been shown to bind FN. At
least a dozen FN-binding MSCRAMMs have been
identified and their corresponding genes have been
sequenced ŽTable 1.. These MSCRAMMs exhibit
structural features typical of other cell wall-anchored
proteins of gram-positive bacteria. At the N-terminus
is a putative signal sequence involved in transport of
the proteins through the cytoplasmic membrane. The
C-terminus is composed of: Ži. a conserved LPXTG
motif which is required for accurate sorting and an-
choring of the proteins to the cell wall; Žii. a hy-
drophobic membrane-spanning region; and Žiii. a tail
of positively-charged residues which remain in the
cytoplasm. Once the protein has been transported to
the cell surface, the LPXTG motif is cleaved between
the Thr and Gly residues. The carboxyl group of the
Thr residue is then linked to a free amino group of a
branch peptide within the peptidoglycan cell wall
ŽSchneewind et al., 1995; Ton-Hat et al., 1997.. The
enzyme responsible for catalyzing these reactions has
not been identified but has been named ‘sortase’. A
proline-rich stretch of residues, thought to span the
peptidoglycan layer of the cell wall, is commonly
found on the N-terminal side of the LPXTG motif in
these proteins.
The prototype FN-binding MSCRAMM, FnbpA
Žalso called FnbA. from S. aureus, has a molecular
mass of approximately 100 kDa. FN binds primarily to
a region adjacent to the proline-rich wall-spanning
region of the MSCRAMM ŽFig. 1A.. This binding
region is composed of a ; 45 amino acid long unit
which is tandemly repeated three times ŽD1, D2, D3
units., followed by a single incomplete repeat unit
ŽD4 unit.. A fifth unit ŽDu. is located approximately
100 amino acid residues N-terminal of D1. FN can
bind to each of the D units, as demonstrated using
synthetic peptide mimics of the sequences. Amino
 D. Joh et al. r Matrix Biology 18 (1999) 211᎐223 213

Table 1
FN-binding MSCRAMMs from staphylococci and streptococci.

Species Protein References

Staphylococcus spp.
S. aureus FnbpA & FnbpB ¨ et al. Ž1989., Jonsson
Signas ¨ et al. Ž1991.

Streptococcus spp.
S. saprophyticus Hemaglutinin Hell et al. Ž1998.

S. pyogenes SfbIrProtein F1 Hanski Caparon Ž1992., Talay et al. Ž1993.

SfbII Kreikemeyer et al. Ž1995.
Protein F2 Jaffe et al. Ž1996.
Opacity factor ŽSof 22. Rakonjac et al. Ž1995.
Glyderaldehyde-3- Pancholi and Fischetti Ž1992.
phosphate dehydrogenase
FBP54 Courtney et al. Ž1994.
M1 Cue et al. Ž1998.

S. dysgalactiae FnbA &FnbB Lindgren et al. Ž1992.

S. equi FNZ Lindmark et al. Ž1996.

S. equisimilis FnB Lindgren et al. Ž1994.

S. mitis Sugano et al. Ž1997.

S. gordonii McNab et al. Ž1994, 1996.

S. pneumoniae van der Flier et al. Ž1995.

S. agalactiae Zabel et al. Ž1996.

S. milleri Willcox et al. Ž1995.

S. anginosus Willcox et al. Ž1995.

Group G streptococci GfbA Kline et al. Ž1996.

acid sequence comparison of the different D units
shows that the D1᎐D4 units are highly conserved
whereas the Du unit is more divergent ŽFig. 1A..
The overall arrangement of the ligand-binding re-
gion in FnbpA is also found in the staphylococcal
protein FnbpB and the streptococcal proteins
SfbIrprotein F1, SfbII, Sof22, protein F2, FnbA,
FnbB, and FNZ, albeit with slight variations ŽFig. 1A..
Furthermore, the sequences of the tandemly repeated
FN-binding units are quite highly conserved among
these proteins ŽFig. 1B. ŽHanski and Caparon, 1992;
McGavin et al., 1993; Lindgren et al., 1994; Talay et
al., 1994; Kreikemeyer et al., 1995; Jaffe et al., 1996;
Ozeri et al., 1996.. In a comparative study, we ana-
lyzed the inhibitory activities of recombinant proteins
derived from the ligand-binding sites of FnbpA of S.
aureus, FnbA and FnbB of Streptococcus dysgalactiae,
and SfbI of Streptococcus pyogenes ŽJoh et al., 1994..
Each of the polyhistidine-tagged proteins inhibited
the interaction of FN with S. aureus, S. pyogenes and
S. dysgalactiae. The cross-inhibitory activities of these
recombinant FN-binding units suggest that the dif-
ferent MSCRAMMs may operate by a similar mecha-
nism and may bind to similar or overlapping regions
in FN. A kinetic analysis of the interaction between
the recombinant binding domains and intact FN, us-
ing BIAcore analysis, indicated that the K d ranged
from 3 to 16 nM ŽHouse-Pompeo et al., 1996.. In
addition, extensive analysis of the residues involved in
ligand binding suggested that a conserved core se-
quence, EDŽTrS.Ž9 or 10 Xs.GGŽ3 or 4 Xs.ŽIrV.DF,
is important for the ligand-binding activity of these
units ŽMcGavin et al., 1991, 1993.. In FNZ, the FN-
binding MSCRAMM of Streptococcus equi subspp.
zooepidemicus, a strong ligand-binding activity was
found associated with the sequence LAGESGET
which is located between the first ŽR1. and second
ŽR2. repeat units of the MSCRAMM ŽLindmark et
al., 1996.. This sequence is also present in the FN-
binding sequence ŽUR or UFBD. located upstream of
the tandem repeats in protein F1 of S. pyogenes Žsee
below. ŽOzeri et al., 1996..
The primary binding site in FN for FnbpA of S.
aureus is located in the 29-kDa N-terminal proteolytic
214 D. Joh et al. r Matrix Biology 18 (1999) 211᎐223

Fig. 1. ŽA. Domain organization of FnbpA from Staphylococcus aureus, the prototype of the FN-binding MSCRAMMs from staphylococci and
streptococci. S, signal peptide; A, non-repetitive region; ␦, two almost identical repeats of 30 amino acid residues with unknown function;
Du-D4, FN-binding units; W, cell wall-spanning region; M, cytoplasmic membrane-spanning region; C, positively-charged intracellular
residues. The cell wall-anchoring LPXTG motif is indicated between regions W and M. ŽB. Alignment of the amino acid sequences of the
ligand-binding units from FnbpA of S. aureus, and FnbA and FnbB of Streptococcus dysgalactiae. Conserved residues that have been
implicated as being important for FN binding are shown in bold face.

fragment of FN ŽFN-N29. ŽMosher and Proctor, 1980..
This fragment is composed of five type I modules,
designated 1FI, 2FI, 3FI, 4FI and 5FI, each of which
is folded tightly through hydrophobic packing and two
disulfide bonds ŽPotts and Campbell, 1994.. The
repetitive nature of the binding sites in both FN and
the MSCRAMM represents a two-dimensional com-
plexity in the interaction of these proteins. This raises
several intriguing questions concerning the binding
mechanism that investigators have attempted to an-
swer. Experiments with recombinant FN fragments,
expressed in baculovirus-infected insect cells, sug-
gested that deletion of any of the five FI modules
abolished the binding of the recombinant proteins to
S. aureus ŽSottile et al., 1991.. Analysis of the interac-
tion of FI modules with FnbpA fragments, using
fluorescence polarization and affinity chromatogra-
phy, revealed that the 4FI᎐5FI pair was the dominant
binding site for the D3 unit of this protein ŽHuff et
al., 1994.. We recently confirmed these findings and
further analyzed the binding patterns of the other D
units ŽJoh et al., 1998.. Each of the full-length D units
ŽDu, D1, D2, and D3., expressed as GST fusion
proteins, recognized the 4FI᎐5FI pair. Additional
binding to 2FI᎐3FI was observed for recombinant D1,
whereas recombinant D4 only bound to 2FI᎐3FI.
Analysis of the FN-binding units from other
MSCRAMMs revealed different binding patterns ŽJoh
et al., 1998.. The B3 unit of S. dysgalactiae FnbB, for
example, bound to 1FI᎐2FI and 2FI᎐3FI, whereas the
A2 unit of S. dysgalactiae FnbA bound only to 4FI᎐5FI
ŽJoh et al., 1998.. According to Huff et al. Ž1994., the
stoichiometry of the interaction of FN-N29 with a
recombinant protein containing the D1᎐D3 units of
FnbpA was 1.9:1. However, the data regarding the
specificity of the individual FI modules and the lig-
and-binding units should be interpreted with caution.
These experiments were performed using short re-
combinant proteins or synthetic peptides correspond-
ing to the MSCRAMM ligand-binding units or the FI
module pairs. How the units and modules interact in
the context of the full-length proteins may not be
directly inferred from experiments with truncated re-
combinant proteins.
Other regions of FN may also interact with
MSCRAMMs ŽKuusela et al., 1984, 1985; Bozzini et
 D. Joh et al. r Matrix Biology 18 (1999) 211᎐223
al., 1992; Sakata et al., 1994.. Protein F1 from
S.pyogenes and GfbA from group G streptococci, for
example, possess a FN-binding unit designated UR in
addition to the repeat unit region ŽOzeri et al., 1996..
The binding site in FN for UR is located in the
40-kDa collagen-binding domain, which is adjacent to
the N-terminal 29-kDa region. The C-terminal hep-
arin-binding domain of FN may also bind to the
streptococcal and staphylococcal MSCRAMMs, al-
though comparatively weakly ŽKuusela et al., 1984;
Bozzini et al., 1992.. The binding site for this domain
appears to lie outside the repeat units in these pro-
teins ŽBozzini et al., 1992.. It is interesting to note
that the collagen- and heparin-binding domains of FN
also contain FI module pairs. However, the
MSCRAMM binding sites in these domains have not
yet been localized.
3. Mechanism of FN-MSCRAMM interaction: insights
from immunological studies
Analysis of the interactions of polyclonal and
monoclonal antibodies with the FN-binding
MSCRAMMs of staphylococci and streptococci has
provided important insights into the mechanism of
ligand-MSCRAMM binding. Interestingly, it has been
noted by several investigators that antibodies raised
against the FN-binding units of FnbpA of S. aureus
do not effectively inhibit the binding of FN to the
MSCRAMM ŽSchennings et al., 1993; Mamo et al.,
1994, 1995.. In a recent study, sera were collected
from more than 30 individuals who had a history of S.
aureus infection. The IgG fractions were isolated and
analyzed for reactivity with FnbpA ŽCasolini et al.,
1998.. Essentially all of the IgG preparations reacted
with the MSCRAMM, consistent with the presence of
the gene encoding FnbpA in ) 90% of S. aureus
clinical isolates ŽMinhas et al., 1995.. Epitope map-
ping, using recombinant truncates of FnbpA, revealed
that the different IgG preparations reacted preferen-
tially with the ligand-binding site ŽDu-D4., suggesting
that this is the immunodominant region of the
MSCRAMM. However, the IgG preparations did not
effectively inhibit the binding of FN to the
MSCRAMM ŽCasolini et al., 1998.. Similarly, we have
found that antibodies raised against recombinant or
synthetic peptide forms of the FN-binding units failed
to display any significant inhibitory activity. In fact, we
observed several monoclonal antibodies ŽmAbs.,
raised against FnbpA of S. aureus and FnbA of S.dys-
galactiae, which actually enhanced the ligand-binding
activities of the MSCRAMMs ŽSpeziale et al., 1996,
manuscript in preparation.. For example, one mAb,
3A10, was incapable of binding FnbA by itself and
only bound to the MSCRAMM if soluble FN was
215
included in the assay. This antibody was shown to
recognize an epitope that was formed in the
MSCRAMM upon binding to FN. Subsequent epitope
mapping experiments, using truncates of FnbA, re-
vealed that the epitope for 3A10 resided within Au,
one of the FN-binding units of the MSCRAMM ŽFig.
1.. Furthermore, 3A10 enhanced, rather than inhib-
ited, the interaction between FN and the Au unit.
The unexpected behavior of these antibodies has
provided invaluable clues as to how the MSCRAMMs
interact with FN. It appears that the FN-binding
repeat units of the MSCRAMMs have a largely unor-
ganized structure and only assume an ordered struc-
ture upon binding to FN. This hypothesis is supported
by the results of biophysical studies of the
MSCRAMM᎐FN interaction ŽHouse-Pompeo et al.,
1996; Penkett et al., 1998.. For example, circular
dichroism spectroscopy of recombinant proteins cor-
responding to the ligand-binding domains of FnbpA,
FnbA, FnbB and SfbI has demonstrated that these
proteins lack a secondary structure and undergo a
structural change upon binding FN ŽHouse-Pompeo
et al., 1996.. Furthermore, gel permeation chromatog-
raphy indicated that the complex of recombinant
D1᎐D3 and FN-N29 was more compact than free
D1᎐D3 Žunpublished observations.. MAbs that react
with ligand-induced binding sites ŽLIBS., such as
3A10, appear to recognize a conformation of the
MSCRAMM induced upon binding to FN. The ability
of 3A10 to enhance the MSCRAMM᎐ligand interac-
tion may be the consequence of the mAb stabilizing
the MSCRAMM᎐FN complex. The binding mecha-
nism of 3A10 is reminiscent of some anti-␣ II b ␤ 3
integrin mAbs which have been reported to bind the
integrin only when it is bound to fibrinogen and which
also enhance fibrinogen binding ŽFrelinger et al., 1990,
1991.. Thus, these anti-MSCRAMM and anti-integrin
antibodies both appear to bind to cell surface ‘recep-
tors’ which undergo conformational changes upon
ligand binding.
The presence of anti-LIBS antibodies in immunized
animals or infected humans is, perhaps, not surpris-
ing. When a FN-binding MSCRAMM is used as an
immunogen, it is likely that the protein quickly inter-
acts with plasma FN before the immune system has
time to recognize the unoccupied form of the binding
site in the MSCRAMM. However, Sun et al. Ž1997. by
immunizing with short subsegments of the FN-binding
units of FnbpA, were able to generate antibodies that
blocked the binding of the individual binding units to
FN . The rationale behind this approach was that, as
the FnbpA-derived peptides were too short to bind
FN, antibodies raised against them should recognize
the binding units in their unoccupied form in the
native protein and thus block the MSCRAMM᎐FN
interaction. The success of this approach is consistent
216 D. Joh et al. r Matrix Biology 18 (1999) 211᎐223
with the idea that rapid binding of the MSCRAMM
to plasma FN hampers the formation of inhibitory
antibodies specific for the unoccupied binding site of
the MSCRAMM. Inhibitory antibodies have also been
successfully raised in rabbits against SfbI of S. pyo-
genes ŽMolinari et al., 1997.. In this study, recombi-
nant SfbI fragments, containing the FN-binding re-
peat units and also the upstream ‘FN-binding spacer
2’ region, were used as immunogens. The latter region
is equivalent to the UR unit Župstream FN-binding
sequence. of protein F1 of S. pyogenes, which appears
to be the dominant FN-binding unit in this
MSCRAMM ŽOzeri et al., 1996.. As the UR unit
binds to the 40-kDa collagen-binding domain of FN,
the ligand-induced-conformation mechanism de-
scribed above may not apply to either the SfbI-FN or
protein F1᎐FN interactions. Thus, it is possible that
the inhibitory activities of the antibodies described by
Molinari et al. may be due to antibodies specific for
the upstream FN-binding spacer 2 region of SfbI.
4. FN-binding MSCRAMMs from other bacteria
A variety of other important pathogenic bacteria,
including Mycobacterium spp., Escherichia coli, and
Borrelia burgdorferi, are also capable of binding speci-
fically to FN. The MSCRAMMs involved in these
interactions have been characterized to varying de-
grees ŽTable 2.. As a detailed account of all these
FN-binding organisms is beyond the scope of this
article, we have chosen to discuss only the interac-
tions that have been analyzed in some detail.
Mycobacterium bo¨ is BCG and Mycobacterium
paratuberculosis bind soluble FN in a saturable, dose-
and time-dependent fashion ŽAslanzadeh et al., 1989;
Valentin-Weigand and Moriarty, 1992.. The number
of FN binding sites was determined to be 8000᎐15 000
and 1600 per bacterium for M. bo¨ is BCG and M.
paratuberculosis, respectively. In addition, Scatchard
analysis indicated the presence of a homogeneous
population of FN-binding components, which bound
the ligand with a K d of 1.25= 10y9 M in the case of
M. paratuberculosis. Several mycobacterial species se-
crete a set of FN-binding proteins Ž30᎐31 kDa. into
the culture medium in ¨ itro. These proteins are known
as the Antigen ŽAg. 85 complex Žrelated to MPB or
MPT51. ŽAbou-Zeid et al., 1988, 1991; Pessolani and
Brennan, 1992; Thole et al., 1992; Ohara et al., 1995;
Naito et al., 1998.. However, as these proteins are
primarily extracellular, it is unclear whether they me-
diate the adherence of mycobacteria to FN. The Ag85
complex is composed of three proteins, Ag85A
ŽMPT44., B ŽMPT59. and C ŽMPT45., which are
genetically distinct but show extensive immunological
cross-reactivity and amino acid sequence similarity,
both within a particular species and between different
species. Recently, two separate FN-binding sites have
been identified on the M. kansasii Ag85B homologue
which appear to be conserved in the Ag85 proteins
from this and other mycobacterial species ŽNaito et
al., 1998.. Studies with synthetic peptides defined 11
residues as a minimum binding unit on Ag85B. This
peptide could inhibit the binding of FN to Ag85B and
to other Ag85 proteins from different species ŽNaito
et al., 1998.. A sequence composed of six residues
ŽFEWYYQ., which has no homology to other known
bacterial FN-binding proteins, was shown to be criti-
cal for FN binding. The binding site in FN for the
Ag85 complex has not been defined. The interaction
of recombinant Ag85B with FN could be specifically
inhibited by gelatin but not by heparin, tentatively
localizing a binding site to the 40-kDa collagen-bind-
ing domain of FN ŽPeake et al., 1993..
Several other mycobacterial FN-binding proteins,
distinct from the Ag85 complex, have been identified.
A 50-kDa FN-binding protein, FAP, has been purified
from M. ¨ accae culture supernatant ŽRatliff et al.,
1993.. Polyclonal and monoclonal antibodies to this
protein recognized a cell wall component in M. ¨ ac-
cae and several other mycobacterial species, including
M. bo¨ is BCG, M. tuberculosis, M. kansasii and M.
a¨ ium ŽRatliff et al., 1993.. Antibodies to FAP inhib-
ited the binding of several mycobacteria to FN and to
mammalian cells ŽKuroda et al., 1993.. Thus, FAP is a
putative FN-binding MSCRAMM on mycobacteria.
FAP homologues have also been identified in M.
tuberculosis ŽFAP-TB. ŽAbou-Zeid et al., 1991., M.
leprae ŽFAP-L. ŽSchorey et al., 1995. and M. a¨ ium
ŽFAP-A. ŽSchorey et al., 1996.. Purified recombinant
FAP-L bound to plasma FN and the C-terminal hep-
arin-binding chymotryptic fragment of FN. A study
with overlapping synthetic peptides revealed two dis-
tinct FN-binding sites conserved among FAPs from
different mycobacterial species ŽSchorey et al., 1996..
These units possess little sequence similarity with
FN-binding MSCRAMMs from other bacteria.
A large number of Escherichia coli strains from
animal and human sources have been reported to
adhere to FN in solid phase assays and to attach to
¨
fibroblasts via FN ŽFroman et al., 1984; Faris et al.,
1988; Ljungh et al., 1990; Yu et al., 1995.. The binding
of this gram-negative bacterium to FN appears to be
mediated by several types of fimbriae Žpili. on the
bacterial cell surface. Fimbriae are hair-like struc-
tures that are composed of repeating major subunit
proteins, often arranged in an alpha helical array
ŽSmyth et al., 1996.. Minor subunit proteins are pre-
sent at the tip and sometimes along the length of
these organelles and often mediate the binding activi-
ties of these structures. Two distinct sites in FN are
targeted by the E. coli receptors, FN-N29 and the
 D. Joh et al. r Matrix Biology 18 (1999) 211᎐223 217

Table 2
FN-binding MSCRAMMs from other bacterial species

Bacterial Species Protein References

Mycobacterium spp. Ag85A ŽMPT44. Borremans et al. Ž1989., DeWit et al. Ž1990.

Ag85B ŽMPT59. Matsuo et al. Ž1988., Thole et al. Ž1992.

Ag85C ŽMPT45. Abou-Zeid et al. Ž1991., Peake et al. Ž1993.

MPT51 ŽMPB. Ohara et al. Ž1995, 1997.

FAP ŽFAP-TB, FAP-L, FAP-A. Abou-Zeid et al. Ž1991., Kuroda et al. Ž1993., Schorey et al. Ž1995, 1996.

Escherichia coli P fimbriae ŽFsoE, FsoF. Westerlund et al. Ž1989, 1991.

Type I fimbriae Sokurenko et al. Ž1992.
Curli Olsen et al. Ž1989.

Campylobacter jejuni CadF Konkel et al. Ž1997.

Flagellin Moser et al. Ž1997.

Salmonella enteritidis Fimbriae Collinson et al. Ž1993.

Yersinia spp. YadA Tertti et al. Ž1992., Schulze-Koops et al. Ž1993.

Porphyromonas gingi¨ alis 150-kDa protein Lantz et al. Ž1991.

Fimbrillin Murakami et al. Ž1996.

Aeromonas salmonicida A-protein Doig et al. Ž1992.

Aeromonas hydrophila Ascencio et al. Ž1991.

Haemophilus ducreyi Abeck et al. Ž1992.

Propionibacterium acnes 80-kDa protein Yu et al. Ž1997.

Fusobacterium nucleatum Winkler et al. Ž1987., Babu et al. Ž1995.

Neisseria gonorrhoeae OpaA van Putten et al. Ž1998.

Neisseria meningitidis Eberhard et al. Ž1998.

Mycoplasma penetrans 65-kDa protein Giron et al. Ž1996.

Treponema pallidum P1 and P2 proteins Peterson et al. Ž1987.

Treponema denticola 53- and 72-kDa proteins and Umemoto et al. Ž1993.
38-kDa axial flagellar protein

Borrelia burgdorferi Fn-BA Probert and Johnson Ž1998.

BBK32 Grab et al. Ž1998.

Borrelia garinii 147-kDa protein Kopp et al. Ž1995.

¨
120-kDa C-terminal region ŽFroman et al., 1984; Faris
et al., 1988; Westerlund et al., 1989; Visai et al.,
1991.. The P-fimbriae ŽPap pili. of uropathogenic E.
coli mediate adherence to immobilized intact FN and
also to FN-N29 and the 120-kDa fragment of FN, but
do not bind soluble FN ŽWesterlund et al., 1989.. The
FsoE and FsoF Žalso called PapE and PapF, respec-
tively. minor subunit proteins of the P-fimbriae have
been implicated in this interaction ŽWesterlund et al.,
1991.. Although it has not been demonstrated that
these subunit proteins bind specifically to FN-N29, it
is noteworthy that they do contain sequences similar
to the FN-binding repeat units of the staphylococcal
and streptococcal FN-binding MSCRAMMs ŽWest-
erlund and Korhonen, 1993..
Campylobacter jejuni, another gram-negative
pathogenic bacterium which causes gastrointestinal
diseases such as diarrhea, can adhere to ECM pro-
218 D. Joh et al. r Matrix Biology 18 (1999) 211᎐223
teins including FN ŽKuusela et al., 1989; Moser and
Schroder, 1995.. Proteinase K treatment of the bacte-
ria abolished FN binding and demonstrated that the
binding components on the cell surface are proteina-
ceous in nature ŽMoser and Schroder, 1995, 1997.. A
major outer membrane protein, CadF, has been iden-
tified as a FN-binding protein. The corresponding
gene has been cloned, sequenced and identified in all
isolates tested so far ŽKonkel et al., 1997.. Another
major outer membrane protein, flagellin, has also
been shown to bind FN and intestinal epithelial cells
ŽMoser et al., 1997.. The sites in the FN molecule
targeted by these putative MSCRAMMs have not
been identified.
The adherence of Porphyromonas gingi¨ alis to ECM
components, such as FN, appears to play a role in the
pathogenesis of periodontal disease caused by this
organism ŽWinkler et al., 1987.. The binding of solu-
ble FN to P. gingi¨ alis was demonstrated to be time-
dependent, specific, reversible, and saturable. The K d
for the interaction was estimated to be in the order of
10y7 M, with approximately 3500 binding sites per
cell ŽLantz et al., 1991.. Lantz et al. identified a
FN-binding protein of 150-kDa in SDS-solubilized P.
gingi¨ alis. Fimbrillin, a major component of the fim-
briae of P. gingi¨ alis, has also been identified to be a
FN-binding MSCRAMM ŽMurakami et al., 1996.. The
fimbriae appeared to interact with the N-terminal
heparin-binding and C-terminal cell-binding domains
in FN ŽMurakami et al., 1996.. In addition, by using a
series of synthetic 20 mer fimbrillin peptides, two
distant regions in this protein have been implicated in
binding FN ŽSojar et al., 1995.. Interestingly, Kontani
et al. Ž1997. recently demonstrated that an arginine-
specific protease expressed by P. gingi¨ alis enhanced
the binding of purified fimbriae to immobilized FN
using BIAcore analysis . It was proposed that the P.
gingi¨ alis protease exposes a cryptic binding site
Žcryptitope. in FN, allowing the bacteria to adhere
more efficiently to FN-coated surfaces.
The syphilis-causing spirochete, Treponema pal-
lidum, has been reported to bind to cultured mam-
malian cells via the C-terminal cell-binding domain of
FN ŽThomas et al., 1985.. The binding of intact FN
and its cell-binding domain to the spirochetes was
saturable, with an estimated K d of 10y7 M. Intact FN
and FN fragments containing the cell-binding peptide,
RGDS, also supported the tip-mediated adhesion of
the human oral pathogen T. denticola ŽDawson and
Ellen, 1990.. These spirochetes express surface pro-
teins of 53 and 72 kDa as well as a 38-kDa axial
flagellar protein that have been shown to bind to FN
ŽUmemoto et al., 1993.. Several reports have indi-
cated that FN may play a role in the interaction of
Borrelia burgdorferi and Borrelia garinii, both causative
agents of lyme disease, with cells and the ECM.
Szczepanski et al. Ž1990. demonstrated that anti-FN
antiserum could inhibit the binding of B. burgdorferi
to a subendothelial matrix.. Recently, a surface-ex-
pressed protein ŽBBK32. with FN-binding activity was
purified from B. burgdorferi and the corresponding
gene was cloned ŽProbert and Johnson, 1998.. The
binding site for BBK32 was localized to the
gelatinrcollagen-binding domain in FN. The ability of
recombinant BBK32 to bind FN and inhibit the at-
tachment of B. burgdorferi to FN was demonstrated
ŽProbert and Johnson, 1998.. A 52-kDa protein ŽFn-
BA. has also been identified as a FN-binding compo-
nent on the surface of B. burgdorferi, however, the
relationship between this protein and BBK32 is un-
clear ŽGrab et al., 1998.. It has been observed that B.
garinii can bind FN in a specific and saturable fashion
and Scatchard analysis revealed the presence of two
types of receptors on the spirochetal surface, one with
high and one with low affinity for FN ŽKopp et al.,
1995.. A 147-kDa protein with high FN-binding activ-
ity was subsequently solubilized from the surface of
B. garinii. This protein, soluble FN and anti-FN anti-
bodies could competitively inhibit the adherence of
B. garinii to epithelial cells ŽKopp et al., 1995..
5. Role of FN-binding MSCRAMMs in bacterial
invasion of mammalian cells
The ability to invade non-phagocytic mammalian
cells may provide bacteria with a niche in which they
are protected from host defense mechanisms or an-
timicrobial agents, which often operate in the extra-
cellular melieu. This may enable the bacteria to per-
sist in the host and may also provide a route for tissue
invasion from a primary site of colonization. Recent
observations suggest that FN-binding MSCRAMMs,
host integrins and FN play important roles in cellular
invasion by a variety of bacteria.
Several gram-positive bacteria, such as Streptococ-
cus spp. and S. aureus efficiently multiply in the
extracellular environment but also have the ability to
invade host cells ŽHamill et al., 1986; Boschwitz and
Timoney, 1994; Lapenta et al., 1994; Almeida et al.,
1996; Calvinho and Oliver, 1998; Winram et al., 1998..
Expression of the FN-binding MSCRAMMs SfbI or
protein F1 promoted the invasion of cultured human
epithelial cells by S. pyogenes and soluble forms of
these proteins inhibited this process ŽMolinari et al.,
1997; Jadoun et al., 1998; Okada et al., 1998..
Polystyrene beads coated with SfbI or protein F1 were
also effectively internalized by these cells ŽMolinari et
al., 1997; Ozeri et al., 1998.. Anti-FN antibodies com-
pletely abolished protein F1-mediated invasion,
whereas increasing FN concentrations resulted in a
significant enhancement of bacterial uptake, suggest-
 D. Joh et al. r Matrix Biology 18 (1999) 211᎐223
ing a role for FN in this process ŽJadoun et al., 1998..
It has recently been demonstrated that the host cell
receptors involved in protein F1-mediated entry of S.
pyogenes are the FN-binding a v ␤ 3 and ␤ 1 integrins
ŽOzeri et al., 1998.. Antibodies against these integrins
effectively inhibited invasion by this bacterium ŽOzeri
et al., 1998.. The entry of S. pyogenes into human
epithelial cells also can be mediated by protein M1, a
multifunctional surface protein, in an FN-dependent
manner which is inhibited by anti-␣ 5␤ 1 integrin anti-
bodies ŽCue et al., 1998.. Our preliminary observa-
tions indicate that the FN-binding MSCRAMMs are
essential for S. aureus invasion of mammalian cells.
Simultaneous inactivation of the genes encoding Fn-
bpA and FnbpB resulted in a 100-fold reduction in
the invasion of HeLa cells by the mutant staphylococ-
cal cells. In addition, soluble fragments of FnbpA and
a RGD-containing peptide inhibited the invasion
Žmanuscript in preparation.. Taken together, these
studies suggest that FN may act as a bridging molecule
between FN-binding bacterial MSCRAMMs and
mammalian cell integrins.
Mycobacteria are obligate intracellular pathogens
which naturally infect macrophages but can also in-
vade non-phagocytic cells such as HEp-2 cells and
Schwann cells ŽBermudez et al., 1995; Schorey et al.,
1995.. The ability to bind FN appears to play an
important role in cellular invasion by these bacteria.
Schorey et al. Ž1995.. observed that the invasion of
Schwann cells was enhanced by pretreatment of the
mycobacteria with FN and that this enhancement was
abolished by anti-FN antibodies or antibodies against
the mycobacterial FN-binding protein FAP.
FN also mediates efficient entry of the facultative
intracellular bacterium Neisseria gonorrhoeae into
HEp-2 cells Žvan Putten et al., 1998.. The FN-induced
entry of OpaA-expressing N. gonorrhoeae, which also
appeared to be dependent on glycosaminoglycans, was
inhibited by RGD-containing peptides and anti-␣ 5 ␤ 1
integrin antibodies Žvan Putten et al., 1998.. The
N-terminal region of FN is believed to interact with
OpaA, through the bridging action of glycosaminogly-
cans, while the type III modules of FN bind ␤ 1
integrins on the HEp-2 cell surface Žvan Putten et al.,
1998.. However, studies have revealed that vitronectin
and ␣ v integrins may be more important for the
internalization of N. gonorrhoeae by other cell lines
ŽDuensing and van Putten, 1997; Gomez-Duarte et
al., 1997; Dehio et al., 1998; Duensing and van Put-
ten, 1998..
The mechanism by which FN-binding MSCRAMMs
trigger the internalization of bacteria by mammalian
cells is unknown. Many bacterial surface proteins that
mediate invasion interact with integrins via bridging
molecules such as FN, or directly bind to integrins, as
has been demonstrated by Isberg and colleagues for
219
the invasin protein of Yersinia spp. ŽIsberg et al.,
1987; Isberg and Leong, 1990; Isberg and Van Nhieu,
1994.. This suggests that these host receptors may
play a central role in the internalization process. The
binding of an integrin to immobilized FN initiates a
complex cascade of cell signaling that leads to reorga-
nization of cytoskeletal components, resulting in cell
spreading and the formation of adhesion plaques.
Engagement of integrins with FN bound to a bacterial
cell surface may trigger a similar process but, due to
the small size and curvature of the bacterial cells, the
interaction may result in internalization of the bacte-
ria. Consistent with this notion, FN-coated polystyrene
beads are effectively internalized by BHK cells ŽMc-
Abee and Grinnell, 1983.. Furthermore, inhibitors of
actin polymerization, such as cytochalasin D, inhibit
the invasion of several bacterial species, suggesting
that the process does involve rearrangement of cy-
toskeletal components of the host cell ŽYoung et al.,
1992; Almeida and Oliver, 1995; Greco et al., 1995;
Menzies and Kourteva, 1998.. How similar are the
intracellular signaling pathways that induce mam-
malian cells to spread on FN-coated surfaces or to
internalize FN-coated bacteria? Does the integrin-de-
pendent internalization of various bacterial species
proceed through similar or different mechanisms?
These are some of the intriguing questions that war-
rant further investigation.
6. Are FN-binding MSCRAMMs virulence factors?
Despite the general belief that FN-binding
MSCRAMMs are pathogenic determinants of mi-
croorganisms, the importance of these MSCRAMMs
in the infection process remains to be determined in
many cases. For example, it is clear from studies with
isogenic mutants lacking FnbpA and FnbpB that these
proteins are responsible for mediating the attachment
of S. aureus to immobilized FN in ¨ itro and contribute
to the adherence of this bacterium to plasma clots
and ex vivo biomaterial ŽVaudaux et al., 1993; Greene
et al., 1995; Vaudaux et al., 1995.. However, data on
the role of these FN-binding MSCRAMMs in animal
models of staphylococcal endocarditis are conflicting
ŽKuypers and Proctor, 1989; Flock et al., 1996; Greene
et al., 1996.. These conflicting results may be due to
the difficulties inherently associated with animal stud-
ies, such as variability among the animals used in
each case. In addition, the outcomes of experimental
infections may be dependent on the animal model
employed. For example, if a collagen-binding
MSCRAMM is responsible for colonization of carti-
laginous tissues, such as joints, it may be more rele-
vant to use an animal model of septic arthritis than a
model of endocarditis to assess the role of this
220 D. Joh et al. r Matrix Biology 18 (1999) 211᎐223
MSCRAMM in virulence. Furthermore, it is likely
that multiple adhesins are involved in different stages
of a particular infection and that a degree of redun-
dancy exists in the activities of these adhesins. All of
these issues should be taken into consideration when
investigating the potential roles of individual
MSCRAMMs in the infection process.
In this review, we have tried to describe the multi-
tude of interactions which exist between bacterial
MSCRAMMs and FN which promote both bacterial
adherence to host tissues and invasion of host cells.
During the course of evolution, pathogenic microbes
have developed elaborate ways to effectively colonize
their hosts, evade host defense systems, interfere with
normal cellular activities and destroy host tissues,
resulting in the symptoms of disease. The expression
of FN-binding MSCRAMMs is only one example of
this adaptation. With the emergence of multiple drug
resistance, it is critical to expand our knowledge of
the molecular mechanisms of microbial pathogenesis
to target the development of novel preventive and
therapeutic strategies.
Acknowledgements
The studies performed in our laboratories were
supported by National Institutes of Health grant
AI20624, by Inhibitex, Inc., and by the Fondo d’Ateneo
per la Ricerca, Italy.
References
Abeck, D., Johnson, A.P., Mensing, H., 1992. Binding of Hae-
mophilus ducreyi to extracellular matrix proteins. Microbiol.
Pathog. 13, 81᎐84.
Abou-Zeid, C., Ratliff, T.L., Wiker, H.G., Horboe, M., Bennedson,
J., Rook, G.A., 1988. Characterization of fibronectin-binding
antigens released by Mycobacterium tuberculosis and Mycobac-
terium bo¨ is BCG. Infect. Immun. 56, 3046᎐3051.
Abou-Zeid, C., Garbe, T., Lathigra, R. et al., 1991. Genetic and
immunological analysis of Mycobacterium tuberculosis fi-
bronectin-binding proteins. Infect. Immun. 59, 2712᎐2718.
Almeida, R.A., Oliver, S.P., 1995. Invasion of bovine mammary
epithelial cells by Streptococcus dysgalactiae. J. Dairy Sci. 78,
1310᎐1317.
Almeida, R.A., Matthews, K.R., Cifrian, E., Guidry, A.J., Oliver,
S.P., 1996. Staphylococcus aureus invasion of bovine mammary
epithelial cells. J. Dairy Sci. 79, 1021᎐1026.
Ascencio, F., Ljungh, A., Wadstrom, ¨ T., 1991. New lectins and
other putative adhesins in Aeromonas hydrophila. Experientia
47, 414᎐416.
Aslanzadeh, J., Brwon, E.J., Quillin, S.P., Ritchey, J.K., Ratliff,
T.L., 1989. Characterization of soluble fibronectin binding to
Bacille Calmette-Guerin. J. Gen. Microbiol. 135, 2735᎐2745.
Babu, J.P., Dean, J.W., Pabst, M.J., 1995. Attachment of Fusobac-
terium nucleatum to fibronectin immobilized on gingival epithe-
lial cells or glass coverslips. J. Periodontol. 66, 285᎐290.
Bermudez, L.E., Shelton, K., Young, L.S., 1995. Comparison of the
ability of Mycobacterium a¨ ium, M. smegmatis and M. tuberculo-
sis to invade and replicate within HEp-2 epithelial cells. Tuber.
Lung Dis. 76, 240᎐247.
Borremans, J., DeWit, L., Volckaert, G., Ooms, J., DeBruyn, J.,
1989. Cloning, sequencing determination and expression of a
32-kilodalton-protein gene of Mycobacterium tuberculosis. Infect.
Immun. 57, 3123᎐3130.
Boschwitz, J.S., Timoney, J.F., 1994. Characterization of the an-
tiphagocytic activity of equine fibrinogen for Streptococcus equi
subsp. equi. Microb. Pathog. 17, 121᎐129.
Bozzini, S., Visai, L., Pignatti, P., Petersen, T.E., Speziale, P., 1992.
Multiple binding sites in fibronectin and the staphylococcal
fibronectin receptor. Eur. J. Biochem. 207, 327᎐333.
Calvinho, L.F., Oliver, S.P., 1998. Invasion and persistence of
Streptococcus dysgalactiae within bovine mammary epithelial cells.
J. Dairy Sci. 81, 678᎐686.
Casolini, F., Visai, L., Joh, D. et al., 1998. Antibody response to
fibronectin-binding MSCRAMM in patients diagnosed with
Staphylococcus aureus infection. Infect. Immun. 66, 5433᎐5442.
Collinson, S.K., Doig, P.C., Doran, J.L., Clouthier, S., Trust, T.J.,
Kay, W.W., 1993. Thin, aggregative fimbriae mediate binding of
Salmonella enteritidis to fibronectin. J. Bacteriol. 175, 12᎐18.
Courtney, H.S., Li, Y., Dale, J.B., Hasty, D.L., 1994. Cloning,
sequencing, and expression of a fibronectinrfibrinogen-binding
protein from group A streptococci. Infect. Immun. 62, 3937᎐3946.
Cue, D., Dombek, P.E., Lam, H., Cleary, P.P., 1998. Streptococcus
pyogenes serotype M1 encodes multiple pathways for entry into
human epithelial cells. Infect. Immun. 66, 4593᎐4601.
Dawson, J.R., Ellen, R.P., 1990. Tip-orientated adherence of Tre-
ponema denticola to fibronectin. Infect. Immun. 58, 3924᎐3928.
Dehio, M., Gomez-Duarte, O.G., Dehio, C., Meyer, T.F., 1998.
Vitronectin-dependent invasion of epithelial cells by Neisseria
gonorrhoeae involves alphaŽv. integrin receptors. FEBS Lett. 424,
84᎐88.
DeWit, L., DeLaCuvellerie, A., Ooms, J., Content, J., 1990. Nu-
cleotide sequence of the 32 kDa-protein gene Žantigen 85A. of
Mycobacterium bo¨ is BCG. Nuc. Acids Res. 18, 3995.
Doig, P., Emody, L., Trust, T.J., 1992. Binding of laminin and
fibronectin by the trypsin-resistant major structural domain of
the crystalline virulence surface array protein of Aeromonas
salmonicida. J. Biol. Chem. 267, 43᎐49.
Duensing, T.D., van Putten, J.P., 1997. Vitronectin mediates inter-
nalization of Neisseria gonorrhoeae by Chinese hamster ovary
cells. Infect. Immun. 65, 964᎐970.
Duensing, T.D., van Putten, J.P., 1998. Vitronectin binds to the
gonococcal adhesin OpaA through a glycosaminoglycan molecu-
lar bridge. Biochem. J. 334, 133᎐139.
Eberhard, T., Virkola, R., Korhnen, T., Kronvall, G., Ullberg, M.,
1998. Binding to human extracellular matrix by Neisseria menin-
gitidis. Infect. Immun. 66, 1791᎐1794.
¨
Faris, A., Froman, ¨¨ M., 1988. Adhesion of
G., Switalski, L., Hook,
enterotoxigenic ŽETEC. and bovine mastitis Escherichia coli
strains to rat embryonic fibroblasts: role of amino-terminal do-
main of fibronectin in bacterial adhesion. Microbiol. Immun. 32,
1᎐13.
¨
Flock, J.-I., Froman, ¨
G., Jonsson, K. et al., 1987. Cloning and
expression of the gene for a fibronectin-binding protein from
Staphylococcus aureus. EMBO J. 6, 2351᎐2357.
Flock, J.-I., Hienz, S.A., Heimdahl, A., Schennings, T., 1996. Recon-
sideration of the role of fibronectin binding in endocarditis
caused by Staphylococcus aureus. Infect. Immun. 64, 1876᎐1878.
Frelinger, A.L., Cohen, I., Plow, E.F. et al., 1990. Selective inhibi-
tion of integrin function by antibodies specific for ligand-oc-
cupied receptor conformers. J. Biol. Chem. 265, 6346᎐6352.
Frelinger, A.L., Du, X., Plow, E.F., Ginsberg, M.H., 1991. Monoclo-
nal antibodies to ligand-occupied conformers of integrin a IIb ␤ 3
ŽGlycoprotein IIb᎐IIIa. alter receptor affinity, specificity, and
function. J. Biol. Chem. 266, 17106᎐17111.
 D. Joh et al. r Matrix Biology 18 (1999) 211᎐223
¨
Froman, G., Switalski, L.M., Faris, A., Wadstrom,¨ T., Hook,
¨¨ M.,
1984. Binding of Escherichia coli to fibronectin. A mechanism of
tissue adherence. J. Biol. Chem. 259, 14899᎐14905.
¨
Froman, ¨¨ M., 1987. Isolation
G., Switalski, L.M., Speziale, P., Hook,
and characterization of a fibronectin receptor from Staphylococ-
cus aureus. J. Biol. Chem. 262, 6564᎐6571.
Giron, J.A., Lange, M., Baseman, J.B., 1996. Adherence, fi-
bronectin binding, and induction of cytoskeleton reorganization
in cultured human cells by Mycoplasma penetrans. Infect. Im-
mun. 64, 197᎐208.
Gomez-Duarte, O.G., Dehio, M., Guzman, C.A., Chhatwal, G.S.,
Dehio, C., Meyer, T.F., 1997. Binding of vitronectin to Opa-ex-
pressing Neisseria gonorrhoeae mediates invasion of HeLa cells.
Infect. Immun. 65, 3857᎐3866.
Grab, D.J., Givens, C., Kennedy, R., 1998. Fibronectin-binding
activity in Borrelia burgdorferi. Biochim. Biophys. Acta. 1407,
135᎐145.
Greco, R., De Martino, L., Donnarumma, G., Conte, M.P., Seganti,
L., Valenti, P., 1995. Invasion of cultured human cells by Strepto-
coccus pyogenes. Res. Microbiol. 146, 551᎐560.
Greene, C., McDevitt, D., Francois, P., Vaudaux, P.E., Lew, D.P.,
Foster, T.J., 1995. Adhesion properties of mutants of Staphylo-
coccus aureus defective in fibronectin-binding proteins and stud-
ies on the expression of fnb genes. Mol. Microbiol. 17,
1143᎐1152.
Greene, C., Vaudaux, P.E., Francois, P., McDevitt, D., Foster, T.J.,
1996. A low-fibronectin-binding mutant of Staphylococcusaureus
879R4S has Tn918 inserted into its single fnb gene. Microbi-
ology 142, 2153᎐2160.
Hamill, R.A., Vann, J.M., Proctor, R.A., 1986. Phagocytosis of
Staphylococcus aureus by cultured bovine endothelial cells: a
model for postadherence events in endovascular infections. In-
fect. Immun. 54, 833᎐836.
Hanski, E., Caparon, M., 1992. Protein F, a fibronectin-binding
protein, is an adhesin of the group A streptococcus Streptococ-
cus pyogenes. Proc. Natl. Acad. Sci. USA 89, 6172᎐6176.
Hanson, M.S., Cassatt, D.R., Guo, B.P. et al., 1998. Active and
passive immunity against Borrelia burgdorferi decorin binding
protein A ŽDbpA. protects aginat infection. Infect. Immun. 66,
2143᎐2153.
Hell, W., Meyer, H.W., Gatermann, S.G., 1998. Cloning of aas, a
gene encoding a Staphylococcus saprophyticus surface protein
with adhesive and autolytic properties. Mol. Microbiol. 29,
871᎐881.
House-Pompeo, K., Xu, Y., Joh, D., Speziale, P., Hook, ¨¨ M., 1996.
Conformational changes in the fibronectin binding MSCRAMMs
are induced by ligand binding. J. Biol. Chem. 271, 1379᎐1384.
Huff, S., Matusuka, Y.V., McGavin, M.J., Inhgam, K.C., 1994.
Interaction of N-terminal fragments of fibronectin with synthetic
and recombinant D motifs from the binding protein of Staphylo-
coccus aureus studied using fluorescence anisotropy. J. Biol.
Chem. 269, 15563᎐15570.
Isberg, R.R., Leong, J.M., 1990. Multiple beta 1 chain integrins are
receptors for invasin, a protein that promotes bacterial penetra-
tion into mammalian cells. Cell 60, 861᎐871.
Isberg, R.R., Van Nhieu, G.T., 1994. Binding and internalization of
microorganisms by integrin receptors. Trends Microbiol. 2,
10᎐14.
Isberg, R.R., Voorhis, D.L., Falkow, S., 1987. Identification of
invasin: a protein that allows enteric bacteria to penetrate
cultured mammalian cells. Cell 50, 769᎐778.
Jadoun, J., Burnstein, E., Skutelsky, E., Hanksi, E., Sela, S., 1998.
Protein F1 is required for efficient entry of Streptococcus pyo-
genes into cultured epithelial cells. J. Infect. Dis. 178, 147᎐158.
Jaffe, J., Natansonyaron, S., Caparon, M.G., Hanski, E., 1996.
Protein F2, a novel fibronectin-binding protein from Streptococ-
221
cus pyogenes, possesses two binding domains. Mol. Microbiol. 21,
373᎐384.
Joh, H.J., House-Pompeo, K., Patti, J.M., Gurusiddappa, S., Hook, ¨¨
M., 1994. Fibronectin receptors from Gram-positive bacteria:
Comparison of active sites. Biochemistry 33, 6086᎐6092.
¨¨ M., 1998.
Joh, D., Speziale, P., Gurusiddappa, S., Manor, J., Hook,
Multiple specificities of the staphylococcal and streptococcal
fibronectin-binding MSCRAMMs. Eur. J. Biochem. 258,
897᎐905.
¨
Jonsson, ¨ C., Muller, H.P., Lindberg, M., 1991. Two
K., Signas,
different genes encode fibronectin binding proteins in Staphylo-
coccus aureus: The complete nucleotide sequence and character-
ization of the second gene. Eur. J. Biochem. 202, 1041᎐1048.
Kline, J.B., Xu, S.H., Bisno, A.L., Collins, C.M., 1996. Identification
of a fibronectin-binding protein ŽGfbA. in pathogenic group G
streptococci. Infect Immun. 64, 2122᎐2129.
Konkel, M.E., Garvis, S.G., Tipton, S.L., Anderson, D.E., Cieplak,
W., 1997. Identification and molecular cloning of a gene encod-
ing a fibronectin-binding protein ŽCadF. from Campylobacter
jejuni. Mol. Microbiol. 24, 953᎐963.
Kontani, M., Kimura, S., Nakagawa, I., Hamada, S., 1997. Adher-
ence of Porphyromonas gingi¨ alis to matrix proteins via a fim-
brial cryptic receptor exposed by its own arginine-specific pro-
tease. Mol. Microbiol. 24, 1179᎐1187.
Kopp, P.A., Schmitt, M., Wellensiek, H.J., Blobel, H., 1995. Isola-
tion and characterization of fibronectin-binding sites of Borrelia
garinii N34. Infect. Immun. 63, 3804᎐3808.
Kreikemeyer, B., Talay, S.R., Chhatwal, G.S., 1995. Characteriza-
tion of novel fibronectin-binding surface protein in group A
streptococci. Mol. Microbiol. 17, 137᎐145.
Kuroda, K., Brown, E.J., Telle, W.B., Russell, D.G., Ratliff, T.L.,
1993. Characterization of the internalization of Bacillus
Calmette-Guerin by human bladder tumor cells. J. Clin. Invest.
91, 69᎐76.
Kuusela, P., Vartio, T., Vuento, M., Myhre, E.B., 1984. Binding
sites for streptococci and staphylococci in fibronectin. Infect.
Immun. 45, 433᎐436.
Kuusela, P., Vartio, T., Vuento, M., Myhre, E.B., 1985. Attachment
of staphylococci and streptococci on fibronectin, fibronectin
fragments, and fibrinogen bound to a solid phase. Infect. Im-
mun. 50, 77᎐81.
Kuusela, P., Moran, A.P., Vartio, T., Kosunen, T.U., 1989. Interac-
tion of Campylobacter jejuni with extracellular matrix compo-
nents. Biochim. Biophys. Acta. 993, 297᎐300.
Kuusela, P., 1978. Fibronectin binds to Staphylococcus aureus. Na-
ture 276, 718᎐720.
Kuypers, J.M., Proctor, R.A., 1989. Reduced adherence to trauma-
tized rat heart valves by a low-fibronectin-binding mutant of
Staphylococcus aureus. Infect. Immun. 57, 2306᎐2312.
Lantz, M., Allen, R.D., Duck, W., Blume, J.L., Switalski, L.M.,
¨¨ M., 1991. Identification of Porphyromonas gingi¨ alis com-
Hook,
ponents that mediate its interactions with fibronectin. J. Bacte-
riol. 173, 4263᎐4270.
Lapenta, D., Rubens, C., Chi, E., Cleary, P.P., 1994. Group A
streptococci efficiently invade human respiratory epithelial cells.
Proc. Natl. Acad. Sci. USA 91, 12115᎐12119.
¨ C., Gurusiddappa, S., Hook,
Lindgren, P.E., McGavin, M.J., Signas, ¨¨
M., Lindberg, M., 1992. Cloning and expression of two different
genes from Streptococcus dysgalactiae encoding fibronectin re-
ceptors. J. Biol. Chem. 267, 1924᎐1931.
¨ C., Rantamaki, L., Lindberg, M., 1994. A
Lindgren, P.E., Signas,
fibronectin-binding protein from Streptococcus equisimilis: char-
acterization of the gene and identification of the binding do-
main. Vet. Microbiol. 41, 235᎐247.
Lindmark, H., Jacobsson, K., Frykberg, L., Guss, B., 1996. Fi-
bronectin-binding protein of Streptococcus equi subsp. Zooepi-
demicus. Infect. Immun. 64, 3993᎐3999.
222 D. Joh et al. r Matrix Biology 18 (1999) 211᎐223
Ljungh, A., Emody, L., Steinruck, H. et al., 1990. Fibronectin,
vitronectin, and collagen binding to Escherichia coli of intestinal
and extraintestinal origin. Zentralbl. Bakteriol. 274, 126᎐134.
Mamo, W., Jonsson, P., Flock, J.-I. et al., 1994. Vaccination against
Staphylococcus aureus mastitis: immunological response of mice
vaccinated with fibronectin-binding protein ŽFnBP-A. to chal-
lenge with S. aureus. Vaccine 12, 988᎐992.
Mamo, W., Jonsson, P., Muller,¨ H.P., 1995. Opsonization of Staphy-
lococcus aureus with a fibronectin-binding protein antiserum
induces protection in mice. Microbiol. Pathog. 19, 49᎐55.
Matsuo, K., Yamaguchi, R., Yamazaki, A., Tasaka, H., Yamada, T.,
1988. Cloning and expression of the Mycobacterium bo¨ is BCG
gene for extracellular a antigen. J. Bacteriol. 170, 3847᎐3854.
McAbee, D.D., Grinnell, F., 1983. Fibronectin-mediated binding
and phagocytosis of polystyrene latex beads by baby hamster
kidney cells. J. Cell Biol. 97, 1515᎐1523.
McGavin, M.J., Raucci, G., Gurusiddappa, S., Hook, ¨¨ M., 1991.
Fibronectin binding determinants of the Staphylococcus aureus
fibronectin receptor. J. Biol. Chem. 266, 8343᎐8347.
McGavin, M.J., Gurusiddappa, S., Lindgren, P.-E., Lindberg, M.,
Raucci, G., Hook,¨¨ M., 1993. Fibronectin receptors from Strepto-
coccus dysgalactiae and Staphylococcus aureus. J. Biol. Chem.
268, 23846᎐23953.
McNab, R., Jenkinson, H.F., Loach, D.M., Tannock, G.W., 1994.
Cell-surface-associated polypeptides CshA and CshB of high
molecular mass are colonization determinants in the oral bac-
terium Streptococcus gordonii. Mol. Microbiol. 14, 743᎐754.
McNab, R., Holmes, A.R., Clarke, J.M., Tannock, G.W., Jenkinson,
H.F., 1996. Cell surface polypeptide CshA mediates binding of
Streptococcus gordonii to other oral bacteria and to immobilized
fibronectin. Infect. Immun. 64, 4204᎐4210.
Menzies, B.E., Kourteva, I., 1998. Internalization of Staphylococcus
aureus by endothelial cells induces apoptosis. Infect. Immun. 66,
5994᎐5998.
Minhas, T., Ludlam, H.A., Wilks, M., Tabaqchali, S., 1995. Detec-
tion by PCR and analysis of the distribution of a fibronectin-bi-
nding protein gene Ž fnb. among staphylococcal isolates. J. Med.
Microbiol. 42, 96᎐101.
Molinari, G., Talay, S.R., Valentin-Weigand, P., Rohde, M., Chhat-
wal, G.S., 1997. The fibronectin-binding protein of Streptococcus
pyogenes, SfbI is involved in the internalization of group A
streptococci by epithelial cells. Infect. Immun. 65, 1357᎐1363.
Moser, I., Schroder, W., 1995. Binding of outer membrane prepara-
tions of Campylobacter jejuni to INT 457 cell membranes and
extracellular matrix proteins. Med. Microbiol. Immunol. 184,
147᎐153.
Moser, I., Schroder, W., 1997. Hydrophobic characterization of
thermophilic Campylobacter species and adhesion to INT 407
cell membranes and fibronectin. Microbiol. Pathog. 22, 155᎐164.
Moser, I., Schroeder, W., Salnikow, J., 1997. Campylobacter jejuni
major outer membrane protein and a 59-kDa protein are in-
volved in binding to fibronectin and INT 407 cell membranes.
FEMS Microbiol. Lett. 157, 233᎐238.
Mosher, D., Proctor, R.A., 1980. Binding and factor XIII-mediated
cross-linking of a 27-kilodalton fragment of fibronectin to
Staphylococcus aureus. Science 209, 927᎐929.
Murakami, Y., Iwahashi, H., Yasuda, H. et al., 1996. Porphyromo-
nas gingi¨ alis fimbrillin is one of the fibronectin-binding proteins.
Infect. Immun. 64, 2571᎐2576.
Naito, M., Ohara, N., Matsumoto, S., Yamada, T., 1998. The novel
fibronectin-binding motif and key residues of mycobacteria. J.
Biol. Chem. 273, 2905᎐2909.
Nilsson, I.M., Patti, J.M., Bremell, T., Hook, ¨¨ M., Tarkowski, A.,
1998. Vaccination with a recombinant fragment of collagen
adhesin provides protection against Staphylococcus aureus-medi-
ated septic death. J. Clin. Invest. 101, 2640᎐2649.
Ohara, N., Kitaura, H., Hotokezaka, H. et al., 1995. Characteriza-
tion of the gene encoding the MPB51, one of the major secreted
protein antigens of Mycobacterium bo¨ is BCG, and identification
of the secreted protein closely related to the fibronectin binding
85 complex. Scand. J. Immunol. 41, 433᎐442.
Ohara, N., Ohara-Wada, N., Kitaura, H., Nishiyama, T., Matsu-
moto, S., Yamada, T., 1997. Analysis of the genes encoding the
antigen 85 complex and MPT51 from Mycobacterium a¨ ium.
Infect. Immun. 65, 3680᎐3685.
Okada, N., Tatsuno, I., Hanski, E., Caparon, M., Sasakawa, C.,
1998. Streptococcus pyogenes protein F promotes invasion of
HeLa cells. Microbiology 144, 3079᎐3086.
Olsen, A., Jonsson, A., Normark, S., 1989. Fibronectin binding
mediated by a novel class of surface organelles on Escherichia
coli. Science 338, 652᎐655.
Ozeri, V., Tovi, A., Burstein, I. et al., 1996. A two-domain mecha-
nism for group A streptococcal adherence through protein F to
the extracellular matrix. EMBO J. 15, 989᎐998.
¨
Ozeri, V., Rosenshine, I., Mosher, D.F., Fassler, R., Hanski, E.,
1998. Role of integrins and fibronectin in the entry of Strepto-
coccus pyogenes into cells via protein F1. Mol. Microbiol. 30,
625᎐637.
Pancholi, V., Fischetti, V.A., 1992. A major surface protein of
group A streptococci is a glyceraldehyde-3-phosphate-dehydro-
genase with multiple binding activity. J. Exp. Med. 176, 415᎐426.
¨¨ M., 1994. Microbial adhesins recognizing extra-
Patti, J.M., Hook,
cellular macromolecules. Curr. Opin. Cell Biol. 6, 752᎐758.
Patti, J.M., Allen, B.L., McGavin, M.J., Hook, ¨¨ M., 1994.
MSCRAMM-mediated adherence of microorganisms to host
tissues. Annu. Rev. Microbiol. 48, 585᎐617.
Peake, P., Gooley, A., Britton, W.J., 1993. Mechanism of interac-
tion of the 85B secreted protein of Mycobacterium bo¨ is with
fibronectin. Infect. Immun. 61, 4828᎐4834.
Penkett, C.J., Redfield, C., Jones, J.A. et al., 1998. Structural and
dynamical characterization of a biologically active unfolded fi-
bronectin-binding protein from Staphylococcus aureus. Biochem-
istry 37, 17054᎐17067.
Pessolani, M.C., Brennan, P.J., 1992. Mycobacterium leprae pro-
duces extracellular homologs of the antigen 85 complex. Infect.
Immun. 60, 4452᎐4459.
Peterson, K., Baseman, J.B., Alderete, J.F., 1987. Molecular cloning
of Treponema pallidum outer envelope fibronectin binding pro-
teins, P1 and P2. Genitourin. Med. 63, 355᎐360.
Potts, J.R., Campbell, I.D., 1994. Fibronectin structure and assem-
bly. Curr. Opin. Cell Biol. 6, 648᎐655.
Probert, W.S., Johnson, B.J.B., 1998. Identification of a 47 kDa
fibronectin-binding protein expressed by Borrelia burgdorferi
isolate B31. Mol. Microbiol. 30, 1003᎐1015.
Rakonjac, J.V., Robbins, J.C., Fischetti, V.A., 1995. DNA sequence
of the serum opacity factor of group A streptococci: identifica-
tion of a fibronectin-binding repeat domain. Infect. Immun. 63,
622᎐631.
Ratliff, T.L., McCarthy, R., Telle, W.B., Brown, E.J., 1993. Purifi-
cation of a mycobacterial adhesin for fibronectin. Infect. Immun.
61, 1889᎐1894.
Sakata, N., Jakab, E., Wadstrom, ¨ T., 1994. Human plasma fi-
bronectin possesses second binding siteŽs. to Staphylococcus
aureus on its C-terminal region. J. Biochem. 115, 843᎐848.
Schennings, T., Heimdahl, A., Coster, K., Flock, J.-I., 1993. Immu-
nization with fibronectin binding protein from Staphylococcus
aureus protects against experimental endocarditis in rats. Mi-
crobiol. Pathog. 15, 227᎐236.
Schneewind, O., Fowler, A., Faull, K.F., 1995. Structure of the cell
wall anchor of surface proteins in Staphylococcus aureus. Science
268, 103᎐106.
Schorey, J.S., Li, Q., McCourt, D.W., 1995. A Mycobacterium leprae
gene encoding a fibronectin binding protein is used for efficient
 D. Joh et al. r Matrix Biology 18 (1999) 211᎐223
invasion of epithelial cells and Schwann cells. Infect. Immun. 63,
2652᎐2657.
Schorey, J.S., Holsti, M.A., Ratliff, T.L., Allen, P.M., Brown, E.J.,
1996. Characterization of the fibronectin-attachment protein of
Mycobacterium a¨ ium reveals a fibronectin-binding motif con-
served among mycobacteria. Mol. Microbiol. 21, 321᎐329.
Schulze-Koops, H., Burkhardt, H., Heesemann, J. et al., 1993.
Outer membrane protein YadA of enteropathogenic Yersiniae
mediates specific binding to cellular but not plasma fibronectin.
Infect. Immun. 61, 2513᎐2519.
¨ C., Raucci, G., Jonsson,
Signas, ¨ K. et al., 1989. Nucleotide sequence
of the gene for a fibronectin-binding protein from Staphylococ-
cus aureus: Use of this peptide sequence in the synthesis of
biologically active peptides. Proc. Natl. Acad. Sci. USA 86,
699᎐703.
Smyth, C.J., Marron, M.B., Twohig, J.M., Smith, S.G., 1996. Fim-
brial adhesins: similarities and variations in structure and bio-
genesis. FEMS Immunol. Med. Microbiol. 16, 127᎐139.
Sojar, H.T., Lee, J.Y., Genco, R.J., 1995. Fibronectin binding do-
main of P. gingi¨ alis fimbriae. Biochem. Biophys. Res. Commun.
216, 785᎐792.
Sokurenko, W.V., Courtney, H.S., Abraham, S.N., Klemm, P., Hsty,
D.L., 1992. Functional heterogeneity of type 1 fimbriae of Es-
cherichia coli. Infect. Immun. 60, 4709᎐4719.
Sottile, J., Schwarzbauer, J., Selegue, J., Mosher, D.F., 1991. Five
type I modules of fibronectin form a functional unit that binds to
fibroblasts and Staphylococcus aureus. J. Biol. Chem. 266,
12840᎐12843.
Speziale, P., Joh, D., Visai, L. et al., 1996. A monoclonal antibody
enhances ligand binding of a fibronectin MSCRAMM Žadhesin.
from Streptococcus dysgalactiae. J. Biol. Chem. 271, 1371᎐1378.
Sugano, N., Tanaka, H., Ito, K., Murai, S., 1997. Arg-Gly-Asp
ŽRGD. peptides inhibit Streptococcus mitis to adhere to fi-
bronectin. J. Nihon Univ. Sch. Dent. 39, 154᎐155.
Sun, Q., Smith, G.M., Zahrda, C., McGavin, M.J., 1997. Indentifi-
cation of D motif epitiopes in Staphylococcusaureus
fibronectin-binding protein for the production of antibody in-
hibitors of fibronectin binding. Infect. Immun. 65, 537᎐543.
Szczepanski, A., Furie, M.B., Benach, J.L., Lane, P.B., Fleit, H.B.,
1990. Interaction between Borrelia burgdorferi and endothelium
in vitro. J. Clin. Invest. 85, 1637᎐1647.
Talay, S.R., Valentin-Weigand, P., Jerstrom, ¨ P.G., Timmis, K.N.,
Chhatwall, G.S., 1993. Fibronectin-binding protein of Streptococ-
cus pyogenes: sequence of the binding domain involved in adher-
ence of streptococci to epithelial cells. Infect. Immun. 60,
3844᎐3847.
Talay, S.R., Valentin-Weigand, P., Timmis, K.N., Chhatwal, G.S.,
1994. Domain structure and conserved epitopes of Sfb protein,
the fibronectin-binding adhesin of Streptococcus pyogenes. Mol.
Microbiol. 13, 531᎐539.
Tertti, R., Skurnik, M., Vartio, T., Kuusela, P., 1992. Adhesion
protein YadA of Yersinia species mediates binding of bacteria to
fibronectin. Infect. Immun. 60, 3021᎐3024.
Thole, J.E.R., Schoningh, R., Janson, A.A.M. et al., 1992. Molecu-
lar and immunological analysis of a fibronectin-binding protein
antigen secreted by Mycobacterium leprae. Mol. Microbiol. 6,
153᎐163.
Thomas, D., Baseman, J.B., Aldrete, J.F., 1985. Fibronectin medi-
ates Treponema pallidum cytoadherence through recognition of
fibronectin cell binding domain. J. Exp. Med. 161, 514᎐525.
Ton-Hat, H., Faull, K.F., Schneewind, O., 1997. Anchor structure of
223
staphylcoccal proteins. A branched peptide that links the car-
boxyl terminus of proteins to the cell wall. J. Biol. Chem. 272,
22285᎐22292.
Umemoto, T., Nakatani, Y., Nakamura, Y., Namikawa, I., 1993.
Fibronectin-binding proteins of a human oral spirochete Tre-
ponema denticola. Microbiol. Immunol. 37, 75᎐78.
Valentin-Weigand, P., Moriarty, K.M., 1992. Protein antigens se-
creted by Mycobacterium paratuberculosis. Zentralbl. Veteri-
narmed. 39, 762᎐766.
van der Flier, M., Chhun, N., Wisemann, T.M., Min, J., McCarthy,
J.B., Tuomanen, E.I., 1995. Adherence of Streptococcus pneumo-
niae to immobilized fibronectin. Infect. Immun. 63, 4317᎐4322.
van Putten, J.P., Duensing, T.D., Cole, R.T., 1998. Entry of OpaA
q gonococci into HEp-2 cells requires concerted action of gly-
cosaminoglycans, fibronectin and integrin receptors. Mol. Mi-
crobiol. 29, 369᎐379.
Vaudaux, P., Pittet, D., Haeberli, A. et al., 1993. Fibronectin is
more active than fibrin or fibrinogen in promoting Staphylococ-
cus aureus adherence to inserted intravascular catheters. J. In-
fect. Dis. 167, 633᎐641.
Vaudaux, P.E., Francois, P., Proctor, R.A. et al., 1995. Use of
adhesion-defective mutants of Staphylococcus aureus to define
the role of specific plasma proteins in promoting bacterial adhe-
sion to canine arteriovenous shunts. Infect. Immun. 63, 585᎐590.
Visai, L., Bozzini, S., Petersen, T.E., Speciale, L., Speziale, P., 1991.
Binding sites in fibronectin for an enterotoxigenic strain of E.
coli B342289c. FEBS Lett. 290, 111᎐114.
Westerlund, B., Korhonen, T.K., 1993. Bacterial proteins binding to
the mammalian extracellular matrix. Mol. Microbiol. 9, 687᎐694.
Westerlund, B., Kuusela, P., Vartio, T., van Die, I., Korhonen, T.K.,
1989. A novel lectin-independent interaction of P fimbriae of
Escherichia coli with immobilized fibronectin. FEBS Lett. 243,
199᎐204.
Westerlund, B., van Die, I., Kramer, C. et al., 1991. Multifunctional
nature of P fimbriae of uropathogenic Escherichia coli: muta-
tions in fsoE and fsoF influence fimbrial binding to renal tubuli
and immobilized fibronectin. Mol. Microbiol. 5, 2965᎐2975.
Willcox, M.D.P., Loo, C.Y., Harty, D.W.S., Knox, K.W., 1995.
Fibronectin binding by Streptococcus milleri group strains and
partial characterization of the fibronectin receptor of Streptococ-
cus anginosus F4. Microbiol. Pathog. 19, 129᎐137.
Winkler, J.R., John, S.R., Kramer, R.H., Hoover, C.I., Murray, P.A.,
1987. Attachment of oral bacteria to a basement-membrane-like
matrix and to purified matrix proteins. Infect. Immun. 55,
2721᎐2726.
Winram, S.B., Jonas, M., Chi, E., Rubens, C.E., 1998. Characteriza-
tion of group B streptococcal invasion of human chorion and
amnion epithelial cells in vitro. Infect. Immun. 66, 4932᎐4941.
Young, V.B., Falkow, S., Schoolnik, G.K., 1992. The invasin protein
of Yersinia enterocolitica: internalization of invasin-bearing bac-
teria by eukaryotic cells is associated with reorganization of the
cytoskeleton. J. Cell. Biol. 116, 197᎐207.
Yu, J.L., Andersson, R., Wang, L.Q., Bengmark, S., Ljungh, A.,
1995. Fibronectin on the surface of biliary drain materials-A role
in bacterial adherence. J. Surg. Res. 59, 596᎐600.
Yu, J.L., Mansson, R., Flock, J.I., Ljungh, A., 1997. Fibronectin
binding by Propionibacterium acnes. FEMS Immunol. Med. Mi-
crobiol. 19, 247᎐253.
Zabel, L.T., Neuer, A., Manncke, B., 1996. Fibronectin binding and
cell surface hydrophobicity contribute to adherence properties
of group B streptococci. Zentralbl. Bakteriol. 285, 35᎐43.