The cytoskeleton

Actin filaments
Structure and organization of actin filaments

An actin monomer is a globular protein with two sites to interact with two other actin monomers→
polymerize to form filaments

​ - Head-to-tail interactions
​ - Each monomer is rotated by 166° in the filaments -> double-stranded helix appearance
​ - distinct polarity
​ - their ends are different
(plus ends and minus ends)

Actin-binding proteins

1. Proteins that stimulate the initiation and elongation of actin filaments

a. Formin, profilin and Arp2/3 complex (actin-related protein)

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 2. Capping proteins that bind to filament ends and filament-stabilizing proteins
a. Tropomyosin → binds along the length of actin filaments
b. Cross-linking proteins:
i. parallel arrays
ii. orthogonal arrays → actin networks

3. Proteins that remodel or modify existing filaments

a. Cofilin → cuts the filaments and generate new ends on them

b. Transient structures →formed in response to

environmental stimuli and involved in cell movement.
c. Pseudopodia → actin filaments cross-linked into a
3D network → Phagocytosis
d. Lamellipodia → sheet like extensions - networks of
actin filaments
e. Filopodia → thin projections of the plasma
membrane supported by actin bundles that extend
from lamellipodia

Cell movement

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Myosin motors

Myosin is the prototype of a molecular motor -> a protein that

converts chemical energy in the form of ATP to mechanical
energy, generating force and movement

Movement In muscle contraction – skeletal and cardiac muscles

Cylindrical bundles of two types of filaments: thick filament of myosin (about 15 run in
diameter) and thin filaments of actin (about 7 nm in diameter). Each myofibril is organized
as a chain of contractile units called sarcomeres. The Z disc defines the ends of the
sarcomere:

I band → only contains thin actin filaments

A band → Contains thick myosin filaments that

overlap actin filaments at the edges of the region

H zone → Middle region with only myosis

Z disc → binds actin filaments by their plus end

Myosin structure

- Large protein (500 kd)

- Two heavy chains →the long alpha helical tails coil around
each other to form dimers

- Two light chains → Associates with the neck of the head

region

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Myosin head groups bind and hydrolyze ATP to provide the energy that drives movement

Nerve impulses stimulate the release of Ca2+ from the sarcoplasmic reticulum

The release of Ca2 ... from the sarcoplasmic reticulum increases the concentration of Ca2 in
the cytosol. Ca+2 is the signal for muscle contraction via the action of two actin filament
binding proteins: tropomyosin and troponin

Troponin is a complex of three polypeptides:

● troponin C (Ca2+-binding)
● troponin I (inhibitory)
● troponin T (tropomyosin-binding)

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Microtubules
Microtubules are hollow rods (varillas huecas) that undergo continual assembly and
disassembly within the cell

Composition: Globulartubulin proteins →

microtubule assembly. Tubulin dimers polymerize to form microtubules

Structure:

- Tubulin dimers polymerize and form protofilaments

- The assemble of 13 linear protofilaments around a hollow core form a microtubule

- Microtubules are polar structures with two distinct ends: a fast-growing plus end and a
slow growing minus end

- β-tubulin binds GTP -> GTP is hydrolysed after polymerization -> the affinity between
dimers decreases -> GDP-tubulin dimer dissociates from the minus end of the microtubule

Dynamic instability → association and dissociation of tubulin subunits from the plus end of
each protofilament. (minus ends are anchored in a Microtubule Organize Centre →
centrosome).

Microtubules grow and shrink depending on the rate of tubulin addition relative to the rate of
GTP hydrolysis.

Microtubules alternate between cycles of growth and shrinkage → Rescue and catastrophe
cycles

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Microtubules-associated proteins

Polymerases → bind to the plus ends of microtubules and accelerate their growth by about
tenfold

Depolymerases → stimulate shrinkage by accelerating the dissociation of GTP/tubulin from

the plus end

CLASP → suppress microtubule catastrophe and promote rescue: stops the disassembly of
microtubules and restarts their assembly

Microtubules stability is regulated by posttranslational modifications of tubulins that provide

them with sites for the binding of specific proteins

Phosphorylation, acetylation, palmitoylation, addition of glutamines or glycines

Interaction with certain proteins in particular localizations – stabilized microtubules -

determine cell shape and polarity –> The TAU protein

The main microtubule Organizing center: The centrosome

It is located adjacent to the nucleus in non dividing cells and contains two centrioles,
oriented perpendicular to each other surrounded by the amorphous pericentriolar material.

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Microtubule motor proteins: Dyneins and Kinesins

Intermediate filaments
​ - Provide mechanical strength to cells
​ - Scaffold for the localization of cellular processes (intracellular signaling)
​ - Composed of a variety of proteins (70 proteins classified in 5
groups)

Type I and II

● Hard keratins – hair and nails

Type III

● Vimentin - organize a network in fibroblasts

● Desmin – muscle cells connect the Z discs

Type IV

● Neurofilaments (NF) light medium and heavy

mature neurons (axons)
● Nestin – Stem cells

Type V

● Nuclear lamina

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Intermediate filaments structure

​ - A central α-helical rod domain (310- 350 aac) - filament assembly

​ - The amino- and carboxy terminal domains (head and tail domains) - determine the
specific functions of the

different intermediate filament proteins

1- The central rod domains of two polypeptide chains are

wound around each other in a coiled-coil structure and form
a dimer

2- dimers of cytoskeletal intermediate filaments associate in

a staggered (forma escalonada) antiparallel fashion to form
tetramers

3- Tetramers assemble end-to-end to form protofilaments

4- Eight protofilaments wound around each other in a

ropelike structure to form a filament

Filaments assembled from antiparallel tetramers, result with equivalent ends →

intermediate filaments are apolar, they do not have distinct plus and minus ends

Phosphorylation regulates their assembly and disassembly within the cell:

- disassembly of the nuclear lamina and breakdown of the nuclear envelope during mitosis

- Vimentin phosphorylation leads to its disassembly and reorganization in dividing or

migrating cells

Intracellular organization of intermediate filaments

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