Microtubule based motor proteins
Two main families of motor protein-kinesin and dynesins are known to
mediate transport along the microtubules.
Kinesin is similar structurally to myosin II in having two heavy chains
and two light chains and two light chain per active motors, these form
two globular head motor domains ad an elongated coiled-coil tail
responsible for heavy chain dimerization.
The yeast has six distinct kinesins, nematode has 16 and humans have
45 kinesins.
There are at least fourteen distinct families in the kinesis superfamily.
Most of them have the motor domain at the N-terminus of the heavy
chain and walk towards the plus end of the microtubule.
Another family has motor domain at the C-terminus and walk in the
opposite direction, towards the minus end of the microtubule.
Members of the kinesin 5 family can self-associate through the tail
domain, forming a bipolar motor that slides oppositely oriented
microtubules past one another, much as a myosin II thick filament does
for actin filaments.
Most kinesin have a binding site in the tail either for a membrane-
enclosed organelle or another microtubule.
Most of the kinesis superfamily members have specific roles in mitotic
and meiotic spindle formation and in chromosomes separation during
cell division.
The dynenins are family of minus-end-directed microtubule motors
unrelated to the kinesin superfamily.
They are composed of two or three heavy chains (that include the
motor and a large and variable number of associated intermediate
chains and light chains.
The dynein family has two branches, the most ancient branch contain
cytoplasmic dyneins, which are typically heavy chain homodimers,
with two large motor domains as head.
Cytoplasmic dyneins are probably found in all eukaryotic cells and
they are important for vesicle trafficking, as well as localization of the
Golgi apparatus near the center of the cell.
Axonemal dyneins, the other large branch include heterodimers and
with two or three motor-domain heads. They are highly specialized for
the rapid and efficient sliding movements of microtubules that drive
the beating of cilia flagella.
Kinesin form a large class of protein family and with diverse function
There are 14 known class of kinesins in animals, protein of the kinesin
superfamily are encoded by about 45 genes in the human genome.
Most of the kinesin are involved in the process, mRNA, organelles ad
chromosomes transport, through the microtubule sliding and
microtubule depolymerization.
As in myosin motors, kinesin families the conserved motor domain is
fused to a variety of class-specific nnmtor domains.
In kinesin-1 has two identical heavy chains and two identical light
chains.
Members of the kinesin-2 family have two different related heavy
chain motor domains and third polypeptide that `associate with the tail
and bind cargo.
Members of the bipolar kinesin-5 family have four heavy chains,
forming bipolar motors that can cross-link antiparallel microtubules
and walking toward the plus end of a microtubule, slide then past each
other.
The inesin-14 motors are only the known class to move towards the
minus end of a microtubule; this class function in mitosis.
Kinesin-13 family has two subunits, but with kinesin domain in the
middle of the polypeptide. They do not have motor activity, but recall
that these are special ATP-hydrolyzing proteins that can enhance
deolymerization of microtubule ends.
Kinesin-1 is a highly processive motor
In kinesin, instead of the rocking of the lever arm, the small movement
of switch loops at the nucleotide-binding site regulate the docking and
undocking of the motor head domain to a long linker region that
connects this motor head at one end to the coiled-coil dimerization
domain at the other end.
When the front (leading) kinesin head is bound to microtubule before
the power stroke, its linker region is relatively unstructured.
ATP-binding of the bound head, the linker region docks along the side
of the heads, this throws the second head forward to a position where
it will be able to bind a new attachment site of the protofilaments, 8nm
closer to the microtubule plus end than the binding site for the first
head.
The nucleotide hydrolysis cycles in the binding site for the first head so
that the cycle of liner docking and undocking allows the two-headed
motor to move in hand-over-hand (head-over-head) stepwise manners
each time taking a discrete 8-nm step.
Kinesin-1 is a dimer of two nucleotide-binding motor domains (head)
that are connected through a long coiled-coil tail.
The two kinesis motor domains work in a coordinated manner, during
a kinesin step the rear head detaches from its tubulin binding sites,
passes the partner motor domain, and then rebinds to the next
available tubulin-binding site.
As the start of each step, one of the two kinesin head, the rear or
lagging head, is tightly bound to the microtubule and to ATP, while
the front or leading head is loosely bound to the microtubule with ADP
in its binding site.
The forward displacement of rear motor domain is driven by an
exchange of ATP for ADP in the front motor domain.
These binding of ATP to this motor domain causes small peptide called
neck linker to shift from a rearward-pointing to the forward pointing
conformation.
This shift pulls the motor domain forward, once it has detached from
the microtubule with ADP bound. The kinesin molecule is now poised
for the next step, which proceeds by an exact repeat of the process.
Dynein Motors transport organelles toward the minus end of the
microtubule
Dynein motor is structurally unrelated myosin and kinesin, but still
follows the general rule of coupling the nucleotide hydrolysis to
microtubule binding and unbinding as well as force generating
conformational change.
This motor protein is very large, consisting of two large (>500KDA),
two intermediate and two small subunits.
It is responsible for ATP dependent retrograde transport of organelles
toward the minus ends of the microtubules in axons, as well as many
other functions.
Cytoplasmic dynein is a two-headed molecule, built around two
identical or nearly identical heavy chains.
Its N-terminal portion forms a tail that binds a set of light chains and
connects to the other heavy chains in dynein molecule, while the major
portion of heavy chain is used to form an elaborate, ring shaped head.
The consists of a planar ring formed from seven domains, six AAA
domains plus the heavy chain C terminal domain, it is therefore a more
complex relative to the hexameric ATPase.
A hook shaped linker region connects the heavy tail to the AAA
domain that is most active as an ATPase.
Between 4th and 5th AAA domains is a heavy chain domain that forms a
long anti-parallel coiled-coil stalk, which protrudes from the structure
and contains the microtubule-binding region.
Before the power stroke N-terminus is attached to the linker that lies
across the AAA domain and associated with the first and 3 rd AAA
domain.
Up o ATP binding and hydrolysis, AAA ring changes conformation
slightly and the linker domain becomes associated with first and 5 th
AAA repeat.
This conformational change rotate the molecule to bring the stem and
stalk closer, resulting in the transport of cargo towards the minus end
of the microtubule.
Cytoplasmic dynein cannot mediate cargo transport by itself.
A large macromolecular assembly often mediates attachment to
membranes.
Cytoplasmic dynein, itself a huge protein complex, required
association with a second large protein complex called dynactin to
translocate organelles effectively.
The dynactin complex includes a short actin like filament that form
from actin-related-protein-1.
Membranes of the Golgi apparatus are coated with proteins ankyrin
and spectrin which are associated with ARP filaments.
The spectrin array probably gives structural stability to the Golgi
membrane and Arp1 mediate attachment of dynein to the organelles.
In other cases, cytoplasmic dynein motors may interact directly with
their cargo.
The cytoplasmic tail of the rhodopsin, the light detecting protein found
in the rod cells of the eye, bind directly to the one of the dynein light
chain and this interaction is required for the normal trafficking of
rhodopsin in the rod cells.
Kinesins and dyneins cooperate in the transport of the organelles through
out the cell
Both Kinesin and dyneins family members play important roles in the
microtubule dependent organization of the organelles in cells (figure).
Orientation of microtubules fixed by the MTOC, the direction of
transport towards or away from the cell center-depends on the motor
proteins.
For example, Golgi apparatus collects in the vicinity of the
centrosomes, where the minus end of the microtubule lies and is
driven there by dynein-dynactin.
In addition secretory cargo emerging from ER is transported to the
Golgi apparatus by dynein-dynactin, conversely the ER spread
through out the cytoplasm is transported by kinesin1
Some organelles of the endocytic pathway are associated with dynein-
dynactin, including late endosomes and lysosomes.
Kinesin have shown to transport mitochondria.
Tubulin modification distinguishes different microtubules and their
accessibility to motors.
The stability and functions of different classes of microtubules are
influenced by post-translational modifications.
For example lysin acetylation, detyrosylation, polyglutamylation and
polyglycylation.
Acetylation of theta-amino group of specific lysine residue and c-
terminus tyrosine can be removed by a carboxypeptidase in alpha
tubulin make microtubule more stable.
The chain of glutamic acid residue is linked to a specific glutamate
residue at the tail region of both alpha and beta tubulin, where a chain
of glycine residues is added to a different glutamic residue and
enhances microtubule stability.
Posttranslational modification of tubulin not only stabilizes the
microtubule but they can also affect the interaction of motor proteins
with them.
