Cytoskeleton

Eucaryotic cells contain protein fibers that are Protein fibers form the cytoskeleton and there are 3 types
involved in of these protein filaments:
- establishing cell shape
- Actin filaments (also called microfilaments)
- providing mechanical strength
- cell movement - Intermediate filaments
- chromosome separation - Microtubules
- intracellular transport of organelles
In addition, a large number of accessory proteins, including
the motor proteins, are required for the properties associated
with each of these filaments

Each type of filament has distinct mechanical properties and

dynamics, but certain fundamental principles are common to
all.
 Actin monomers assemble into long helical polymers with polarity

Some functions of actin filaments are:

- to provide mechanical strength to the cell by forming a band
under the plasma membrane
- link transmembrane proteins to cytoplasmic proteins
- form contractile ring during cytokinesis in animal cells
- cytoplasmic streaming
- generate locomotion in cells such as white blood cells and
amoeba
- Interact with myosin to provide force of muscular contraction
 Actin Filament Assembly

The critical concentration Cc is the concentration of G-actin monomers in equillibrium with

actin filaments.
Monomer concentration below Cc no polymerization occurs.
Monomer concentration above Cc assembly occurs until monomer concentration reaches Cc.
Nucleation is the rate-limiting step in the formation of a cytoskeletal polymer

The time course of actin polymerization in a test tube

 Actin Filaments grow faster at (+) end than at (-) end

The rate of addition of ATP-G-actin is much faster at the (+) end than at the (-) end
(rate of dissociation is similar) -> lower critical concentration (Cc) at (+) end

in steady state -> filament grows preferentially at the (+) end

If actin conc. is between Cc- and Cc+ (steady

state) -> actin subunits flow through
filaments by attaching to (+) end and
dissociating from (-) end
Treadmilling phenomen -> involved in
movement of cells

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Actin regulators: Thymosin and Profilin
 Actin Filament Branching

Nucleation of
branching
mediated by
Arp2/3

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 Myosins - Cellular Motor Proteins

Tail:
-> Locatized to cellular membranes
-> vesicle attached (cargo)

Form thick filaments in muscles

S1 motor domain

Head -> Motor domain (S1) -> ATP depentend

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Myosin heads walk along actin filaments -> towards (+) end

Sliding-filament assay:
Myosin tail absorbed onto glass
surface -> a solution of actin
filaments allowed to flow
through
In presence of ATP myosin
heads walk towards (+) end of
actin filaments -> sliding of
filaments

-> Movement of labeled

actin filaments

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 Myosins – Motor proteins responsible for cell movement

These are the most important 3 myosins

Loss of more specialized ones -> causes deafness/blindness
Skeletal muscle contraction is regulated by Calcium and actin binding proteins

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Myosin motion along actin

Length of the neck domain ->

determines rate of movement

Step size

-> Moves in 72 nm steps

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Actin fibers in the muscle

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Microtubules Arrangement

Flagella

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Microtubules Assembly preferably at (+) end
Nucleation of microtubule assembly

-> Treadmilling

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 Microtubules Assembly/Disassembly

Colchicine and Taxol:

Drugs that interfere with Microtubules
Assembly/Disassambly
Colchicine: 2500 years ago Egyptians treated heart
problems
Nowadys: treatment of gout, skin and joint diseases
Taxol: (stabilizes Microtubules)
Anticancer agents -> treatment of ovarian cancer

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Microtubules Dynamic Instability

Presence of GTP-β-tubulin cap

determines stability

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 The centrosome
Cell polarity including the organization of cell organelles, direction of membrane
trafficking, and orientation of microtubules is determined by microtubule-organizing
centers (MTOCs).
Figure 16-30b Molecular Biology of the Cell (© Garland Science 2008)
 Kinesin-catalysed Vesicle Transport

Carries cargo

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Kinesin-1 uses ATP to walk down a microtubule to the (+) end

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Dynein-catalysed Vesicle Transport

Moves towards the (-) end of microtubules

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Cooperation of Myosin and Kinesin at the cell cortex

Secretory vesicles are handed over

from Kinesin to Myosin -> last part of
secretory pathway

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 Intermediate filaments

• So named because of its diameter in

striated muscles (diameter ~10nm).
• Its core structure is anα-helical coiled coil.
• N- and C-terminal domains vary considerably
in size.

• Inter mediate filaments are flexible but

stable.
• Primary assumed function of intermediate
filaments is to prevent excessive stretching.
• In general, much less is known about
intermediate filaments compared to actin and
microtubule.
 Polymerization modification drugs

Actin
latrunculin Actin monomer Latrunculia
stabilization
phalloidin Actin monomer Amanita
stabilization
cytochalasin Actin filament
stabilization,
end‐capping
Microtubule
taxol Microtubule
filament
binding
colchicine Tubulin sub‐unit Autmn crocus
binding
Vincristine, Tubulin sub‐unit Vinca rosea, Yew
Nocodazole binding tree