Types of vitamins

(a functional view)

Vitamins and blood function III:

Vitamin K
Biochem/NS 510, D. Eide

I.!
II.!

Overview of blood clotting

Vitamin K
Structure
Function- !-carboxyglutamate (GLA) synthesis
The role of GLA in blood clotting
Effects of Vitamin K deficiency
Vitamin K and heart disease

! Vitamins of energy metabolism

acetyl/acyl transfer reactions
redox cofactors
carboxylation
decarboxylations
decarbox, trans- & deaminations

! Vitamins and blood function

one-carbon transfer reactions
carboxylations

Vitamin C
Vitamin E

! Vitamins as signaling molecules

Vitamin A
Vitamin D

Functions of blood

! Excretion
Carry CO2 to lungs- bicarbonate (HCO3-)
Nitrogen (glutamine, urea)
Bilirubin (heme breakdown)
! Respiration- Carry O2 to tissues- hemoglobin, red blood cells
! pH and osmolarity control
! Immunity
B lymphocytes- antibody production
T lymphocytes- antigen recognition, B cell activation, cell killing
Phagocytic cells- macrophages
! Signaling- Hormones, vitamin D
! Hemostasis- coagulation and clotting

folate, B12
Vitamin K

! Vitamins as antioxidants

How does blood clot?

! General- maintain a constant environment (homeostasis)

! Nutrition
glucose, amino acids, lipids (lipoproteins)
vitamins, minerals

pantothenate (B5)
riboflavin (B2), niacin (B3)
biotin
thiamin (B1)
pyridoxine (B6)

Hemostasis
! Primary hemostasis
cellular response
vessel damage exposes collagen
platelets bind to collagen
release factors that activate other platelets
cells form a hemostatic plug- blocks leak
! Secondary hemostasis
protein response- coagulation factors
occurs simultaneously as 1 response
cascade of factors and proteolysis
results in formation of fibrin from fibrinogen
forms threads that strengthen the hemostatic plug
! Two secondary hemostasis cascades
intrinsic pathway
extrinsic pathway
both require vitamin K

Vitamin K

Vitamin K structures
O

Enzyme cofactor
Fat-soluble vitamin (our first)

]3

]6

Not covalently bound (vs. biotin)

Phylloquinone
(plant sources)

Adds CO2 to protein substrates (vs. biotin)

Menaquinone-7
(gut bacteria)

O
Needed by only one enzyme- !-carboxylase
-,

Requires CO2 and O2, not HCO3 ATP

Menadione
(supplements)

Function of Vitamin K
Prothrombin

! VKs only known function:

! adds a 2nd -COOH to GLU in protein.
! one enzyme: !-carboxylase (ER)
! Double -COOH GLU is called GLA
! GLA = !-carboxyglutamic acid
! GLA becomes a Ca2+ binding site.
! Some proteins need bound Ca2+ to work
--- N - C - C - N--

GLU

COO-

CO2, O2,
Vitamin K

--- N - C - C - N-COOCOO-

GLA
!-carboxyglutamate

Why do we need GLA-dependent Ca2+ binding?

Mechanism of VK Action

! Blood clotting (2 hemostasis)

--- N - C - C - N ---

GLU

GLA

+ O2 + CO2

COOOH

OH

--- N - C - C - N ---

COOCOOO

O
VKOR

NADP+
NADPH + H+

Vitamin K
epoxide

! 4 of 13 clotting factors contain GLA residues

! Also 4 proteins that regulate clotting
! K = Koagulant Factor

! Bone remodeling
! Bone GLA protein (osteocalcin)
! Mobilize bone calcium?
! Regulated by Vitamin D

O
R

! Matrix GLA protein

! Function unknown
! Keep Ca soluble?

! Kidney GLA protein

! Regulated by Vitamin D
! May increase Ca2+ resorption

! 11 total, are there others?

What happens in K deficiency?

How do we know whats optimal?

! Severe: You bleed to death.

! Direct measurement of nutrient

! Subclinical: problems with bone minerals?

! VK deficiency thought to be risk factor for osteoporosis
! Not supported by evidence

! Direct assessment of its homeostasis

! easily assayed material (e.g. blood, urine)

! Assessing Deficiency:
! Plasma prothrombin levels
! Clotting assay
! normal K: 11-13 seconds to clot
! deficient K: > 25 seconds to clot

Whos at Risk for low K Status?

! partitioning of bolus, tracer, input/excretion

! Direct measure of biochem actions

! enzymatic rxn, gene expression levels

! Indirect measure of physiological action

! tissue function / strength / differentiation

! Epidemiological relationships
! diet recalls, intake vs. disease, supplement trials

VK-based Anticoagulants

! Newborns - no VK stores

! Impaired gut function/malabsorption

! CF, IBD, pancreatitis, liver disease, bile duct
obstruction, long term total parenteral nutrition

! Chronic Antibiotic Treatment

OO
OH

OH

OH

OH

OH

! Few gut bacteria- little contribution to status

! Breast milk low in vitamin K and low stores due to
poor placental transfer

OH

Dicoumerol

Coumadin/Warfarin

! which wipe out gut sources of VK

! Anticoagulant Therapy
! some anticoagulants block VK action
! Dicoumerol & Coumadin (aka Warfarin)
! Used in treatment of heart disease and stroke

These drugs are VK mimics;

Block recycling of VK after carboxylation rxn.

Warfarin History

Mechanism of VK Action
--- N - C - C - N ---

GLU

--- N - C - C - N ---

GLA

+ O2 + CO2

COOOH

COOCOOO

OH

! Discovered here at UW

O
VKOR

NADP+

Vitamin K
epoxide

NADPH + H+

Warfarin Issues
! ~2 million Americans take warfarin
! Ideal dose varies per individual
! Too much, bleed
! too little, no benefit
! Doctors prescribe by experimentation and monitoring
! Side effect- frequent cause of emergency room visits
! What influences optimal dose?
! Age, height, weight, other medications
! Variations in two genes
! VKORC1
! Encodes VK epoxide reductase
! Differs in activity between people
! CYP2C9
! Cytochrome P450
! Metabolizes warfarin for excretion
! Differs in activity between people

!
!
!
!

K.P. Link, 1940s

Feed causing hemorrhaging in cows
Moldy silage made from sweet clover
Took Harold Campbell 5 yrs to purify 6 mg

! Work supported by WARF

! Still gets $ from its use
! Pays graduate fellowships

O
R

! Effective rat poison- slow acting

! Prescribed as anticoagulant for use in humans 1954
! Dwight Eisenhower treated with it in 1955
! Joseph Stalin poisoned with it in 1953?

! John Suttie discovered how it inhibits VKOR in 1978

! Former chair of the Nutritional Sciences department

! Faculty Senate supports K.P. Link Drive 2008