Cytoskeleton

&
Cell Motility
 cell movements
• If cells want to change their shapes or locations,
such things are needed.
cytoskeleton-- a cytoplasmic network system of protein
fibers
ATP (adenosine triphosphate) -- ATP → ADP+Pi + 1.72KJ
motor proteins-- proteins that convert the energy stored
in ATP into motion
 three types of cytoskeleton
• The cytoskeleton is a network of protein
filaments throughout the cell’s cytoplasm.
• Microfilaments (Actin filaments)
7-8nm

• Intermediate filaments
10nm

• Microtubules
24nm
Muscle contraction
• Sarcomere is the
function unit of
skeletal muscle.

myosin

actin
 Thin filaments are formed from
actin filaments
• Actin monomer is called G-actin.
• Actins are different isoforms in muscle cells from
nonmuscle cells
• After polymerization, G-actins form F-actin .

Actin monomer
with bound ATP Actin filaments have polarity
Monomer — G-Actin
• has a globular
structure, consists
of two lobes 
separated by a actin-ADP

cleft.
• The cleft can bind
Mg2 ＋ & ATP (ADP
while in filament). + actin-ATP

• ATP hydrolysis is
slow for monomer
but very quick for
filament.
 stage of assembly
• Lab experiments ATP-actin polymerizes faster than ADP-actin
show G-actins will ATP-actin dissociates slower than ADP-actin
polymerize to form
F-actin
spontaneously if the
concentration of
ATP, Mg2 ＋ added
into G-actin solution
are high enough.
 Actin filaments have polarity.
F-actin hydrolyzes ATP  ADP.

Treadmill caused by addition match removal

 Thick filaments are formed from
Myosin
• rod-like, contains 1 or 2 globular
head(s), each one can bind MF and
hydrolyze ATP, special tail sections can
carry cargoes.
The mechanism of
myosin movement
Bipolar myosin filaments generate force of
contraction.
 Muscle Contraction
• Tropomyosin binds along the lenth of microfilament,
and overlays the myosin binding sites on microfilament
in resting muscle.

Tropomodulin
caps MF.

Troponin can bind Ca2 ＋ , regulates

muscle contraction.
• Triads are complexes consisting of T tube
flanked on each side by terminal cisternae of
SR.
Calcium triggers contraction of actin and
myosin filaments.
• Ca2+ binds to
troponin C ,
tropomyosin
move away
from the
binding sites
allows myosin
heads to bind,
initiating
muscle contra
ction
.
weak binding
rigor mortis

tight binding

Actually, we can not move even one step.

• smooth muscle cell
doesn’t contain
troponin.
Actin- myosin interactions act in
cytokinesis
Actin- myosin interactions
cause nonmuscle movements

Some types of myosins

transport vesicles and
organelles.
Actin crosslinking proteins generate and
maintain different networks of filaments.

filamin
• In cell, the concentration of G-actin monomers is
more than 100μM, far more than polymerization
needed, but there are still lots of monomers in
cytoplasm, why?

actin-sequestering protein
Actin filaments determine cell
shapes
normal
biconcave disk shape
→ sphere-shape
abnormal
Hereditary spherocytosis, AD
Symptoms include anemia, jaundice, splenomegaly.
 Microtubule, MT
• Microtubules are
hollow cylindrical
tubes.
• Its walls are made up
of 13 protofilaments
which correlate with
one another laterally.
Structure of MT
• Microtubules can form different
structures, including the mitotic
spindle and centriole of dividing cells,
and the core of cilia and flagella.
Structure of MT
• Each protofilament is assembled from protein
subunits of tubulin, which is itself a heterodimer,
consisting of one α-tubulin and one β-tubulin.

• Because each assembly

unit contains two
nonidentical components
(alternating α-tubulin
andβ-tubulin), the
protofilament has
polarity.
αβ-tubulin heterodimer
• Each monomer has a binding site for one molecule
of GTP. Onβ-tubulin, GTP can be hydrolyzed and
produce GDP.
(taxol ,colchicine, Mg2 ＋、 Ca2 ＋ )
γ-tubulin
• γ-tubulin is a critical component
in microtubule nucleation.
 Microtubules polymerization
• MT singlet is labile
structure, can do
rapidly polymerize and
depolymerize.
• in vitro, assembly of
microtubules happens
spontaneously, by
adding tubulins(1mg/ml
), Mg2+, GTP, and EGTA
(which binds Ca2+, an
inhibitor of
polymerization),
incubate at 37℃.
polymerization in vivo
• In an interphase fibroblast cell, a seemingly
haphazard and random network of
microtubules permeates the entire cytosol.
• Unlike the case in vitro, we can see that the
cytosolic microtubules are arranged in a
hub-and-spoke array that lies at the center of
a cell.
Microtubules Organizing Centers
(MTOC)
• The microtubule spokes radiate from a
central site occupied by the centrosome,
which is the primary microtubule-organizing
center (MTOC) in many interphase cells.
•Centrioles are surrounded
by amorphous, electron-
dense pericentriolar material
(PCM)

•The polarity of these microtubules is

always the same: the minus end is
associated with the centrosome, and the
plus (or growing) end is situated at the
opposite tip.
polymerization in vivo
• The minus end is
capped byγ-tubulin
complex ring.
• Only the plus end
can grow or shrink .
 Microtubule Dynamics -- The growth or shrinkage of a
microtubule depends on the hydrolysis speed of GTP
compared with adding speed of Tubulin-GTP.

• A growing microtubule has the open

sheet containing tubulin-GTP subunits.
• When the tube has begun to close,
forcing the hydrolysis of the bound GTP.
• The tube has closed to its end, leaving
only tubulin-GDP subunits. GDP-tubulin
subunits have a curved conformation
compared to their GTP-bound
counterparts, which makes them less
able to fit into a straight protofilament.
• GDP-tubulin subunits at the plus end of
the microtubule is released as the
protofilaments curl outward from the
tubule and undergo catastrophic
shrinkage.
The function of Microtubules
• determine cell’s shape

• form the spindle fibers

to capture & separate
chromosomes
during mitosis
The function of Microtubules
• provide a set of "tracks" for cell
organelles and vesicles to move along.
MAP (microtubule-associated
proteins)
• Several other proteins are associated
with MT.
MAP1, MAP2, Tau, MAP4
Mortor proteins
• Myosin (work with MF)

• Kinesin

• dynein
Kinesins and Dyneins
• 2 heavy chains can bind MT
alternatively and hydrolyze ATP,
special tail sections can carry cargoes.
Kinesins and dyneins mediate
bidirectional transport of organelles
along microtubules.
• Nearly all kinesins move cargo
toward the (+) end of MTs,
Dyneins transport cargo in
opposite direction.

Axonal transport
Kinesins and dyneins mediate
bidirectional transport of organelles
along microtubules.
• Nearly all kinesins move cargo
toward the (+) end of MTs,
Dyneins transport cargo in
opposite direction.
 The function of Microtubules
• cause movement of flagella and cilia.
Basal Bodies work as MTOCs.
movement of flagella and cilia
• Cilia and flagella
have a 9 (doublet) + 2
(singlet) pattern of

microtubules.

Each movement turn, there are

only half of dynein proteins can
be activated.
movement of flagella and cilia
• Cilia and flagella move
when the microtubule
doublets slide past
one another.
movement of flagella and cilia
 Take home Points & Learning
Objectives
Points
• Cytoskeleton, ATP and motor proteins help cells to
maintain the ability of movement.
• Polymerization & depolymerization are essential for
cytoskeleton.
• Cells control the distribution of organelles through
motor proteins and the cytoskeleton.
Objectives
• List examples of how cytoskeleton trigger cell
movement and try to explain the mechanism of the
case.
The end