The Cytoskeleton #2

Molecular Motors
 Motor Proteins
• These proteins bind to polarized Cytoskeletal filaments
and use the energy derived from repeated cycles of ATP
hydrolysis to move steadily along it

• Dozens of different motor proteins coexist in eukaryotic

cells.

• They differ in the type of filament they bind to (actin or

microtubules), the direction they move along the
filaments and the “cargo” they carry
• The cytoskeletal motor proteins associate with
their filament tracks through a head region (or
motor domain) that binds and hydrolyzes ATP

• The motor domain determines the identity of the

track and the direction of movement along it.

• The tail of the motor protein determines the

identity of the cargo
 The Three Groups of Cytoskeletal Motor
Proteins
• Myosins
– Bind actin and moves towards the positive end
(except of myosin VI which moves towards the
negative end)
• Kinesins
– Bind microtubules and moves towards the positive
end
• Dyneins
– Bind microtubules and moves towards the minus end
 The Myosin Superfamily

The first motor protein identified

was skeletal muscle myosin
called myosin II
The Myosin II Bipolar Thick Filament
Direct evidence for the motor activity of the
myosin head
 Sequence comparisons amongst diverse
eukaryotes indicate that there are at least 37
distinct myosin families

• Some mysoins have only been found in plants

(VIII and XI) others only in vertebrates (IX)

• The myosin tails have apparently diversified to

permit the proteins to bind to other subunits and
cargos.

• Yeast contain 5 myosins, C. elegans at least 15

and humans 40.
Myosin Superfamily Members
The exact functions for all mysoins remain
to be determined

• Myosin II associated with contraction in muscle and

non muscle cells
• Myosin V is involved in vesicle and organelle transport
• Myosin I generally involved in intracellular
organization
• 9 human myosins are expressed primarily or
exclusively in the hair cells of the inner ear. Mutations
in 5 of them are known to cause hereditary deafness
 The Kinesin Superfamily
• First identified in the giant axon of the squid.

• Similar structurally to myosin II in having 2 heavy

chains and two light chains per motor

• Like myosin kinesins share a common motor domain

• Yeast have 6, C. elegans 16 and human about 45

kinesins
Some Examples of Kinesins
• 1- C-terminal binds cargo
• 3 – functions as a monomer
and moves membrane
enclosed organelles along
microtubules
• 5 – bipolar and slides
microtubules past each
other
• 13-lost motor activity binds
to microtubules to increase
dynamic instability
• 14-moves towards the
minus end not the plus end
of microtubules
 The Dynein Family
• Cytoplamsic dyneins are
important for vesicular
transport and the
localization of the Golgi

• Axonemal dyneins are

Composed of 2 or 3 heavy chains containing
the motor domain and a large and variable
number of associated intermediate and light
chains
highly specialized for the
rapid sliding of
microtubules that drive the
beating of cilia and flagella

Largest of the molecular motors and among the

fastest
Structural Similarity of Myosin and Kinesin
• The two classes of motor proteins track along different
filaments and have different kinetic properties, and they show
no identifiable amino acid sequence similarities.

• However determination of the 3D structure of the motor

domains of both myosin and kinesin has revealed that these
two domains are built around nearly identical cores.

• The central force-generating element that the two types of

motors proteins have in common include the site of ATP
binding and the machinery necessary to translate ATP
hydrolysis into an allosteric conformational change.
The motor domain of myosin is substantially larger than that of
kinesins, about 850 aa compared to 350
Cycle of structural changes that allows
myosin to walk along an actin filament
Cycle of structural changes that allows
myosin to walk along an actin filament
The Mechanochemical Cycle of Kinesin
• 1 - rear lagging head bound tightly
and leading head loosely bound

• 2 – exchange of ADP for ATP in the

front motor causes a small protein
termed “neck linker” to shift to a
forward pointing conformation

• 3 - This shift pulls the rear motor

forward once it has detached by
the hydrolysis of ATP and Pi release
The Power Stroke of Dynein
• Dynein contains 6 AAA domains 4
of which can bind ATP but only
one retains the major ATPase
activity

• In the ATP bound state the stalk is

detached

• ATP hydrolysis causes the stalk to

attach

• Release of ADP and Pi results in

the power stroke involving
rotation of the head and stalk
relative to the tail domain
 Motor Protein Kinetics are Adapted to Cell
Functions
• A single dimer of kinesin I moves in a highly
progressive manner traveling for hundreds of
ATPase cycles before dissociating
• Whereas mysoin II cannot move progressively and
make just one or two steps before dissociating
• However what mysoin loses in processivity it gains
in speed e.g. linked mysoins can move its filament
20 steps during a cycle time whereas kinesins can
only move 2
 Motor Protein Kinetics are Adapted to Cell
Functions
• Within each class of motor protein movement speed
vary widely, from 0.2 to 60mm in myosins and from
0.02 to 2mm for kinesins

• These differences arise from the fine tuning of the

mechanochemical cycle

• The velocity can be decreased by decreasing the rate

of ATP hydrolysis or by increasing the proportion of
time spent bound to the filament track
• E.g. mysoin V spends up to 90% of its cycle bound to the
filament track compared to 5% for myosin II.
• The change in each step size can be regulated by changing the
length of the lever arm or the angle through which it swings
Motor proteins mediate the movement of
membrane enclosed organelles
• Dynein alongside a large
number of accessory
proteins can associate
with membrane
enclosed organelles.
• Dynactin is a large
complex that can bind
weakly to microtubules,
Dynein itself and the Arp
1 actin related filament
The cytoskeleton can localize specific RNA
molecules to establish cellular
asymmetries
Cells can regulate motor function e.g. the
assembly of non muscle myosin II
Specialized Organization of motor proteins
and filaments e.g. Skeletal Muscle
The Sarcomere
The Sarcomere
Muscle Contraction
The Troponin Complex
The Troponin Complex
Microtubules can form flagellum or cilium
Ciliary Dynein
The Bending of an Axoneme