Cell

biology 6

Cytoskeleton
Microlaments, microtubules and
intermediate laments
i pkry 6 ks or To6ks ct f kcy k i c pn r k nck os C6t s r)
s
obi 6p ps t o rcT Tc
ks as TkTr6p c kcbs o: r
N E E#A 66 i s b r ri n v oho rv
s c rnks r k or
s hocks t s

pkry 6 ks ks ck6r i
6k a ks k i kcbs 66 r s
nckho r i t i os cp kc
i cs rnkc f s i t

c rnks ro 6 kc i r bc ba ks
k i ckt krkt r os k T bi c
66r

nos i os b nc k 66r 66
ohoroks pkyos rorj

nckho 66r f oi t i s o6
rc s bi
 ot c kTs t j or f s i k i i os as F ks o s os b C6t s r s i i o y c
t pkros C6t s rD
ckho s ro6 rc s bi Dt o s Ts aks or k s 6 66r k f oi rs i t i s o 6 rc rr
i k Tcr f i s 66r c rc i
kTbi rs t kr Tc 6 k i i c pn r k pkry 6 6 C6t s r) f i s 66r
c c f oi ks s c r6 rk6Taks r s s ks oks o cb s rvi os ct o
C6t s r rTchoh f i o6 t krk i c rk i pkry 6 ks or rckp D
 kTs os i pkn6r t k t krs ot 6 66r

i p pno 6 6p kct s f kcy i ckTbi kTi pkn6r t vrTcckTs os b i s T 6 Tr s

w s os b kTk i 66n coni cpD
s s i kc k i n6r t t t cs 66x 66gTs aks r rT i r rt krkt r
f i c i w cs 6 k i t t cs or ks s k i k s ki c 66
s ct o C6t s r c 6 rk kTs f oi os i s T 6 Tr( t ri k os ct o
C6t s rvi Ts c6o r s rc s bi s r i s T 6 c s h 6kn
 D ln AD # n Di l #Ac l Tu i n T #T eDMAi D i Dl i T ul TD A

nck os t ks kt c
Ml u l TE ks rorr k s c6 ck
TD l A kt o s f oi b6k T6c
TA# s TA#
T c boks r oi c s D
n ocr k t ks kt cr
r rk o k kct
ot c1 1 rb b c
c t c1 DD

s c6 ck kt o s r k
o c s os ct o
C6t s nck os r) rot o6c
os ro: s r MT s
66pn r kct C6t s r k
rot o6c ot c s
os cs 6 rcT Tc

6k T6c i s o 6c boks r wnkr j ks i rTc k i C6t s vos c f oi ki c

kt nks s r k i pkn6r t ( hc p bc 6p os ki ro: s t os k o r MT s ckt ks
os ct o C6t s nck os k s ki c
 D ln A D A# n Di i Dl D ##i A i Dn A #i Dl i i
Ts s 6 ks b i 6 s bi k s ch 66w ks rvnckho os b rr s a6 os cs 6 c os kc t s
k i r wc t 6p 6ks b s Cs 66 w s roks rD
Ts s os t Tr 6 66r s os noi 6o6 66r rT i r i kr k i ryos D
s 6 6i r 66rv v p rc i os b s orco Tas b i k 6k 6 6p n n6o kc rvy n
66r s i oc t t cs r ckt c y os b os c rnks r k t i s o 6 ri c

i k noi 6o 6 66r or rc i ) s f kcy k os ct o C6t s r s rt krkt 6

gTs aks r i w s r i ckTbi i ri h 6knr s roks s 6ot or i w s k rc i os bD
Ts aks r 6 ks f c nc r s ) T r t g kc kct a ks k i 66r s cTnTc
 j os
noi 6o6 66r
qj s n
os
ks s ah FarrT 66rvt Tr 6
66rvs b6o6 66
Aj os s ch
66r
Gj f i oi
rc s bi s i s T 6 c
t t cs k 6 6 s ot 6 66r

y ca s r c i t kr oh cr 6r r k os ct o C6t s ) i noi 6oTt i r or

kf s oras ah t owTc k y ca s nck os rD n o6 o: y ca s r 6 rk k Tc os i o cv
i crvs 6f r

c as r ks s i ckTbi rt krkt r) 6os b k i obi s ro6 rc s bi Dot nkcs kc

rc i os b k ryos s orco Tas b rc rrD
) os f i o i t Ta ks r os i y ca s b s r os c c
f oi i kct a ks k y ca s C6t s r os i no ct orv h s kT i 6 r k 6or cos b
 rrkcp nck os rvu# N vi ckrrF6os y i C6t s Ts 6 r os k rcks b c cp
T# DT D l E # i T pu l N E# l #L RAn N fe #A c DT n A l TDE E# i eDT
N # n Di e DT i AR i Dl E DEl i A D i n Ti Tn i

A l TDE E# n6 as
Ta ks r os i b s kc n6 as Tr hr a s b i Tt s or r i kt os r Tc r k
no ct k6pror T66kr rot n6 w T r p orcTnaks k ryos y ca s jvt Tr T6c prckni p T r
p orcTnaks k os ct o C6t s r os t Tr 6 jvs s Tck b s c aks T r p
orcTnaks k s TckC6t s rjD o 6 yos b Ts aks 6 n6 as b s o f oi os f p rk
oci
 E # l # n A
i s o 6 rTnnkc kc s , i t s ro kc i ckt a s
ks s s T 6 Tr k s wc 66T6c t cowz

r 6 pkn6r t o os ct o C6 t s r
T s T 6 c 6t os orr r t 6 s c F kct i 66 ohoroks vf i s i sT 6 c s h 6kn
c y r kf s Tcos b t okror s i s c F kct r os i T bi c 66

ah 6 t os r)
t os kn i o r os 6T os b
nckb cor
k 6k pn k
c okt pkn i p) 6 y k
ks f oi as r
ye l N # En l r w- - vv ph, wyf TArw, m
w, - vi l uw- - vv
 orr r t 6p s c r r t 6p k i s T 6 c 6t os c ks ck66 p i ni krni kcp6a ks
s ni krni kcp6a ks k i 6t os r p nck os yos r rD
6 t os r c ni krni kcp6 vi ks r MT s ks kct a ks 6 i s b f y s r i os os b
f s i ct cr s T r r i C6t s k 6 6n c
ni krni kcp6a ks i s k t okror T r r i 6t os r k c r r t 6

t os P s t os
 A l TDE E# i

6ks b s c 6a h 6p ra i k66kf T r k nck os i cn o 6p orr r t 6 os ks 6k a ks s

c r r t 6 os s ki c
t o ckT T6 r bckf kT ckt rt 6 6rcT Tc i
w s kf c i 66n coni cpv c rpr t k c yr f oi os i 66v6 ks b f i o i h ro 6 rv
kcbs 66 rvs ki c 66 kt nks s r c t kh
 o ckT T6 r w s ckt s kcbs o: os b s c
rT i r j s ckrkt v j
rnos 6 nk6 vkc j i r 6 k pk o6oTt D

t 66F p 6
c bT6
r r t 6pj

n ct s s
rcT Tc rj
! To6 ckt rT Ts orO T T6os O
ot c kt nkr k f k h cp
rot o6c b6k T6c nck os r c
c i

! T T6os ot cr r y kb i cFF
kct i f 6 6k i i k66kf
p6os co 6t o ckT T6 D

! T F6oy rcT Tc or t k
A n c6 6 6 i
6os c i o s k T T6os ot cr
f oi Fs FT T6os
6 cs a s b 6 ks b or 6 s bi

! T T6os t k6 T6 r c 6 6
c cs b os i nckkC6t s r
f oi i rt kco s a ks vrk
i i t o ckT T6 i r
rcT Tc 6nk6c op
! FT T6os s vor s
i ki cvi FTFT T6os s vor
i
 E E#A uT#Ln l AS i l Tn E # NT i AD i T Dl Ti Tn D

s ckrkt ks rorr k s t kcni kTr t cowk nck os ks o s os b i sDE E#A cos br i

s T 6 t o ckT T6 bckf i D s s ot 6 66rvi s ckrkt ks o s r no c k s cok6 rv
i t Tn k p6os co 6 c cp k ri kct o ckT T6 rD
n A Ei T n A l TDE E# or t os i s ckrkt vi h os b bckf s ckt
s T 6 a s b cos bvf i c r i u#Ei T n A l TDE E# Ai l A D LDTu# i n
s T 6 a s b ro r nckho p i sDE E#A cos br os i s ckrkt
 l TMA n A l TDE E# i i TM L n A i D A#ADL
i t o ckT T6 C6t s bckf r s
ri cos yr os n s s 6p k or
s obi kcrD
i c cp k t o ckT T6 r s i kc os
s ckrkt or ks as T6 6p i s bos b
r s f t o ckT T6 r bckf s k6
t o ckT T6 r ri cos y

ri cos y nc a6 6p s i s v rT s 6pv
rc bckf os b b o s vkc ot p
orn n c kt n6 6pvk c n6
TcoC c FT T6os s p s f t o ckT T6 ckt i rt
nk6pt co: rnks s kTr6p FT T6os cos b
f i s i obi ks s ca ks v Tos
i 6ohos b 66vi ks s ca ks k D
c FT T6os or s krk i obi
L n A A i D A#ADLer t r
ckt i os cos ro n op k T T6os
t k6 T6 r k i p ck6p:
 i p ck6pror ks ck6r i bckf i k t o ckT T6 rD
j T T6os ot cr c cpos b t e os t kc abi 6p
k ks s ki c i s k T T6os ot cr c cpos b
t eD

j i c kc vt o ckT T6 r i i h c ri 6p
T T6os ot cr i oc s f oi kTs s k
y n bckf os bD

j ckt at k at vi kf h cv rn o6 6p f i s
t o ckT T6 bckf i or r6kf vi rT Ts or os i or
n f o66i p ck6p: i oc k kc c ri
rT Ts or 6k f oi i h at k os D
n or i c p 6kr( i c cpos b
rT Ts or c 6 rr abi 6p kTs os i nk6pt c s
c c o6p c 6 r ckt i c s vrk i i
t o ckT T6 bos r k ri cos y ks as TkTr6p
 A l TDE E# i l n A D A L # T i i n #L
Ai i i n #Lr An uTl D D Tl uT# l AS NT ## ARAi AT
o ckT T6 r s nc h s ckt orr r t 6os b o or n6Tr s or r o6o: p, i t s
k s ki c t k6 T6 kc 66rcT Tc r F nc h s r T T6os nk6pt co: a ks b k
kcbs 66 rPyos k i kc r

i Di Eu A #L Tl AS i Li D n T n A l TDE E# i #A cA i # D u l Di T D ##

66r s c n ADTi Ai n A l TDE E# i kt os oa6 6p t kc ps t o vrf o i os b f s bckf os b

s ri cos yos b t T i t kc c MT s 6p i s pkn6r t o t o ckT T6 r s kct 6 6p kD
i or s 6 r i t k orr r t 6 cn o 6p s i s c r r t 6 os k i t oka rnos 6 Df oi
s f cs kt 6p bckf os b t o ckT T6 r i ks s k yos k i kc r
T# l ADL) i s i of oi 66 kc wi i r os os b nck os ) r 6 ah r o6o: a ks 6 r k
cn o c kco s a ks k i ) ks h ci 66k rcks b6p nk6c o: kct
 vf i o i os r abi 6p k c T T6os s nc h s r or nk6pt co: a ks os k
t o ckT T6 rvi t oka rnos 6 c no 6p orn n c r s i 66r6 6r os i t o 6 k
t okrorvTs 6 k nc aaks or i ckt krkt r os k f k bckTnrD

i r i knnkro aks i t k6 T6c 6 h 6) os r abi 6p k t o ckT T6 r s

nc h s r i t ckt 6kros b rT Ts orD T r s f rT Ts or s ra66 vi
t o ckT T6 r s bckf T s s kri cos yD

TD i i DAi i n r T l Tn Ti Tn i l NT
s kks 6p k r r t 6p T6 rk k orr r t 6p 6 rk MT6 6p ot nkcs
k6 i o os vw k6vhos coras vs hos 6r as vc Tr os i 6os o 6 c t s k s c
 A l TDE E# i Tl AS D D l ATl T D ##
s ot 6 66r c
ch 66rv kc nTkTs w ks ckt ks s k i 66s s co r ckt i ki c ##i
i u A #AS Tl i l NT R D Al T# A uu l DEi uTi ANT DTM l D i AD T i l NT
6 6i t o ckT T6 r os i w ks nkos os i rt oc aks vf oi i oc n6Tr s r kf c i
w ks ct os 6
6ks b i r kco s c yr i 66or 6 k r s t t cs h ro 6 r s nck os r kc
r c aks vi c t os i 66 k p Tc MToc c f p i s k i w ks
ce os ki oc aks r)
i c h cr oc aks
coh s p s ki c r k
n TDTl ul TD A i D
i os f c ce cco r
t co6 r os b r p
i an k i w ks kc
nck T p i
c y kf s k nck os r
s ki c t k6 T6 r
y kf c i 66
k pD
 TDTl l TD A i l AR Dl ##E# l Dl i uTl D
pkn6r t or r s k os ks as T6 t kaks
cb s 66 r t kh os ri kcrc Frkn t s s c) r6 kcp t kh t s
as s t o ckT T6 r t o i or

ho kF s i s ot b r k l , s c k s os h c c s ch 66vs Tt ckTr
t t cs h ro 6 r s t ok i ks co c nc r s Di nckho r Cw ct
k c c s Di r ot b r f c c kc os ch6 r k Gt o66or ks rD

n TR n Di l l D L n TDTl ul TD A i eM A Ei D l L l AR
l Tn l u D L # i T L l T#Li Ai DT Dl R #i D A#L #T D N
# n DTl D n A l TDE E# A i A # Al NT
 TDTl ul TD A i n TR #T n A l TDE E# i Ei A D Al #T E# l i
t kkc nck os r i t kh 6 ks b pkn6r t o t o ckT T6 rr A i A i t kh kf c r n6Tr s
L A ) t kh kf c r i t os Tr s r
cA i A i L A i l TD An l i MAD DMT #T E# l s A A i i A # D A#
kkc nck os f o66 , i k
t o ckT T6 os r c krn oC
t ss c
o 6k t kkc nck os os r r 6p
k rkt 66 kt nks s vrT i r
h ro 6 kc s kcbs 66 s
ct os r i pn k c bk i
i t kkc nck os s cs rnkc

#T E# l i T cA i A
L A l SLn i MAD s
L l T#LSA p i f NRADLD

ckho r i s cbp kc p 6 k ks kct a ks 6 i s b r os i i i s 6 ok t kh

6 ks b i t o ckT T6 p p 6 k os os bvc 6 r vs c os os b k i t o ckT T6
 A i A ) ot c k f k i h p
i o s rv i kt n6 w k 6obi
i o s vf oi k6 t k6 T6c
f obi k A3v
#l #T E# l
T D L #T
DT u Al T i n ##
#T E# l M A
T D A D #A D A i

i i kt o s vf i o i os r
t o ckT T6 r s vor
c rnks ro 6 kc i t kkc ahop
k yos ros

o 6 kt os or c rnks ro 6 kc
os os b k i t t cs k
h ro 6 r t kr6oy 6p i ckTbi i
yos ros 6obi i o s D
Kinesin Super family: 14 known classes of kinesins

Varia#on on the basic structure:

Kinesin1 and 2 involved in
organelle transport. + end
directed motor

Kinesin5 has 4 heavy chains
assembled in a bipolar manner
to interact with an?parallel
microtubules and is a + end
directed motor

Kinesin 13, has motor domain in
the middle of the heavy chain,
no motor ac?vity but destabilize
microtubules
i A # n T# E# T cA i A n TR i #T n A l TDE E#
j 6 cks t o ckbcn i k
ros b6 yos ros t k6 T6
ri kf os b i f k i
kt o s ri
j i c ct rvr nc
p os ch6 r k r ks v
c kc i t kh t s k
s os oho T6 yos ros F
t k6 T6 j 6 ks b
t o ckT T6 j
rn k DA t t Pr

j co r k t k6 T6c
t k 6r k i f k i rk
yos ros t k6 T6 v
ri kf os b i kf i p c
i kTbi k nck rroh 6p
f 6y i oc f p 6 ks b
t o ckT T6 os r co r k
3Fs t r nr ks 6 j
Kinesin-1 uses ATP to walk down a microtubule
Processive: takes hundreds of 8nm steps without
disassocia&ng

Binding of one head to -tubulin subunit induces
loss of ADP
Allowing strong binding of nucleo#de free kinesin
head.
Now forward head binding to ATP leads to
conforma#onal change in the corresponding linker
domain, making it docking to the head and causes
rear head to swing forward and become new
forward head that binds to the next -tubulin
subunit
Now forward head loses ADP, rear head hydrolyzes
ATP, becomes a weakly bound ADP, repea#ng the
cycle again

Dynein motors move organelles towards minus end of microtubules

Large protein Dynein binds to cargo through another

Has 2 large, 2 intermediate and 2 large protein complex: Dynac?n (11
small subunits subunits). Dis?nct cargo binding (ARP1,
Moves towards () end CapZ complex), microtubule binding,
Retrograde transport of organelles dynein binding parts.
 A i A i
L A i TTu l D
DT Dl i uTl D
Tl ## i
D Tl TE TEDD
##
 A l TDE E# i #u DT l l D Tl ## i A Ec l LTN ##
i s 66r c c f oi k6 i o os t o ckT T6 r
kkc nck os r orr r t 6 jv s k6bo i s b i oc 6k a ks
, i k i k6bo i k66n r r k i s c k i 66ks T# Af i o i or
t t cs rvnT66i s k , i k s p ki c kcbs 66 v cb t s r os k rt 6 6
k6bon nc Tr i h ro 6 rvf i o i orn cr i ckTbi kTi pkn6r t D
ki c f p 6 ks b cTb c t kh v i kcb s 66 r c Tcs k i oc kcobos 6
t o ckT T6 rvos f c nkroaks rv cb b p t kkc nck os r t khos b 6 ks b i c F
kf c i 66 s cD kct t o ckT T6 rD
 kr os os b f oi
T T6os ri kf r i
t o ckT T6 r
kcbs o: 66T6c
kcbs 66
orco Taks
Cytoplasmic dyneins
mediate retrograde
transport of
organelles towrds
end (cell centre)

Kinesins mediate
anterograde
transport towards
cell periphery

Organelles have
some type of motor
associated with them
in a cell type specic
manner
The cytoplasm of a eucaryo?c cell is supported and spa?ally organized by a
cytoskeleton of intermediate laments, microtubules, and ac?n laments.
Intermediate laments: stable, ropelike polymers of brous proteins that give
cells mechanical strength. Some types underlie the nuclear membrane to form the
nuclear lamina; others are distributed throughout the cytoplasm.
Microtubules are s?, hollow tubes formed by the polymeriza?on of tubulin
dimer subunits. They are polarized structures with a slowgrowing minus end and a
fast-growing plus end.
Microtubules are nucleated in, and grow out from, organizing centers such as the
centrosome. The minus ends of the microtubules are embedded in the organizing
center.
Many of the microtubules in a cell are in a labile, dynamic state in which they
alternate between a growing state and a shrinking state. These transi?ons, known
as dynamic instability, are controlled by the hydrolysis of GTP bound to tubulin
dimers.

Each tubulin dimer has a ?ghtly bound GTP molecule that is hydrolyzed to
GDP aMer the tubulin has assembled into a microtubule. GTP hydrolysis
reduces the anity of the subunit for its neighbors and decreases the
stability of the polymer, causing it to disassemble.

Microtubules can be stabilized by proteins that capture the plus enda
process that inuences the posi?on of microtubule arrays in a cell. Cells
contain many microtubule-associated proteins that stabilize microtubules,
bind them to other cell components, and harness them for specic func?ons

Kinesins and dyneins are motor proteins that use the energy of ATP
hydrolysis to move unidirec?onally along microtubules. They carry specic
membrane vesicles and other cargoes and in this way help to maintain the
spa?al organiza?on of the cytoplasm.

A l T # n Di
 A l T # n Di AR i i u DT ##i

j t o ckho66o j ks c a6 Ts 6 r os i pkn6r t
j ri 6oy t es Cs b c6oy t enckcTroks r ckt i
6 os b b k t khos b 66
j ks c a6 cos b Tcos b 66 ohoroks
 Ac#n laments are thin,
exible protein threads


each lament may be thought
of as a two stranded helix with
a twist repea?ng every 37 nm

Strong interac?ons between
the two strands prevent the
strands from separa?ng

Branching:
 as C6t s r s bckf
p i oaks k as
t ks kt cr oi c s v
Ti c k bckf i or
r c i n6Tr s
i s i t os Tr s

as t ks kt c c co r
abi 6p kTs v
f i o i or i p ck6p: k
rkks c i
os kcnkca ks k i
as t ks kt c os k i
C6t s D
i p ck6pror k k
os s as C6t s
c T r i rc s bi k
os os b f s
t ks kt cr s
c r r i r o6op k
i nk6pt cD

T 6 ka i p ck6pror i c p nckt k r nk6pt co: a ks vi 6nos b i 66k orr r t 6

C6t s r c i p i h kct
Polymeriza#on of ac#n invivo is ini#ated by proteins that promote lament
forma#on. Can spontaneously assemble invitro
This may require signals.

Formin protects from

capping
So no ATP hydrolysis
and therefore + end
keeps growing
Arp proteins ini#ate branching of ac#n laments whereas formins
promote unbranched laments
 L l TD A i A DT N n T AL Di l Tu l N i
s os
k as t ks kt cr os i
pkrk6vnc h s as b i t
ckt os b k i s rk
as C6t s r

s n
t e ks ck6
as r r t 6p i
h s os b cks k
t obca s b 66

kt i k6 as os nc 6 6 6
Ts 6 rvki cr os b 6F6oy

6rk6os vr h c r C6t s

r rk o f oi t kkc
nck os rt pkros j k kct
ks c a6 Ts 6 rvr os
k s kct c yr 6 ks b f i o i t kkc t Tr 6 66rD
nck os r cs rnkckcbs 66 r
 s i or ncknkr t i s ort kc
## l M#A u i T N 66t kh t s v as
nk6pt co: a ks i 6 os b b
Tl i l D A D N sl A Tl D F k i 66 i n6r t
n TR ## Tl M l t t cs kcf c nckcTroks j
s kct r s f c boks r k as
kc w
f nkos r k s i kcb c t
f s i as C6t s r s
i rTc ks f i o i i 66or
cf 6os b , i t s jD

ks c aks i c c k i 66
i s cf r i k p k i 66
kcf c c aks jD

f s i kcb nkos r c
r 6ori i cks vs k6
ks r c c 6 r i yr
i 66 cf 6r kcf c Di rt
p 6 or c n kh c s kh c
b os vt khos b i 66 kcf c os
r nf or r i oks D
N # n Di ##TM An # ##i DT
n Al D m i t a cf os b k C ck 6r
ri kf os b l , s 6t 66onk o s
Cs C6knk o nckg as b ckt or
rTc v rn o6 6p os i c boks r k
i 6 os b b D
j o 6r k i c cs b t s k
as C6t s r os i c c boks r k
i C ck 6r c ri kf s vf oi
c ckf i r nkos as b
kf c i n6Tr s k i
C6t s rD
s s os b 6 cks t o ckbcn i
ri kf os b 6t 66onk o s C6knk o
i 6 os b b k i Tt s
C ck 6r t obca s b os T6Tc Ds
c ckf ri kf r i oc aks k 66
t kh t s D
M T N # n Di uEi i D # A T
# n ##AuT AEn Tl M l

T 6 a ks k s f as C6t s r
j or t o p cn kt n6 w r
j, i k i ro r k nc woras b
C6t s rDi c rT6as b cs i os b rcT Tc
nTri r i n6r t t t cs kcf c D
i n6Tr s r k i as C6t s r kt
nck p n nos b nck os r jvf i o6
i t os Tr s r k as C6t s r s c c i
s c k i 66 ks as T6 6p orr r t 6
i ckTbi i aks k nk6pt co: os b
nck os r

i f k as r f i k6 i c p
Ts cbk r ks as T6 c c f c t kh t s
T k i r r t 6p k C6t s r i cks
s i oc orr r t 6p i c c D
Myosins Are a Large Superfamily of Mechanochemical Motor Proteins

Three family members myosin I, myosin II, and myosin Vpresent in all
eukaryo#c cells
Plus end motors except for myosin VI that is a minus end motor

Specic ac#vi#es of these myosins dier but all func#on as motor proteins
myosin II powers muscle contrac#on, as well as cytokinesis.

Myosins I and V take part in cytoskeletonmembrane interac#ons, such as
the transport of membrane vesicles.

All myosins consist of one or two heavy chains and several light chains, which
generally have a regulatory func?on

Head, neck, and tail domain organiza?on is found in all myosin heavy chains.
Structure of
myosin II
l Tn n T # i i i T LTi A
LTi A
RL A e
R l A # #A D A i
Tn n n l i #L n LTi A i
A D l D Al D#L i A #
MAD n n l

s 6p 6r r i
r r t 6 r os k
onk6c C6t s r

r row6obi
i o s rvf k
i s
t kh r cbk
Sliding Filament assay used to detect myosin powered movement
 c

pkros i Tr r k nT66ks s as
C6t s

k ) Cct , i t s k as h cp ri kc
6oh r ( cobkTc t kcar???j

os os b )c 6r as
i p ck6prorv s ora66c t o s kTs
i or r or 6 6 - k y 0vi s cbp or rkc
r s 6r a rc i os i t pkros

t pkros os r as vc 6 r r os i
6r a s cbp c 6 r or kTn6 k t khos b as
66 - nkf c rcky .

c t o s r abi 6p kTs r or c 6 r
s s f or
pkros i p ck6pr r ks kc i r n oy r
Op?cal trap determines the step size
and force generated by a single myosin
molecule
 pkros t kh r os
- i s kh c i s . pn
k t kh t s s r n
ro: or 2qs t D
i i r ks i
6 s bi k i 6os y c
6 s bi f r nc o k
6 k ckrr AJ s t
 Myosin powered movements: skeletal muscle contrac#on

Each mul?-nucleated muscle cell has several myobrils consis?ng of ac?n and myosin
laments
Structure of skeletal muscle sarcomere



I bands: ac?n laments

Thin ac?n laments interdigitate with
thicker myosin laments to for A bands



A sliding lament model has
been proposed for contrac?on
Of skeletal muscles
 The arrangement of thick
myosin and thin
ac?n laments in the
relaxed state

In the presence of ATP and
Ca2+, the myosin heads
extending from the thick
laments walk toward the
ends of the thin laments.

Because the thin laments
are anchored at the Z disks
movement of myosin pulls
the ac?n laments toward
the center of the
sarcomere, shortening its
length in the contracted
state

Arrival of the ac?on poten?al triggers the opening of Ca channels

from SR releasing Ca into the cytosol. Ca concn. induces
tropomyosin and troponin that block myosin bing to move away,
allowing myosin interac?on
Ac?n lament if
stabilized by the
accessory
proteins

CapZ caps plus
end
Tropomodulin
caps the minus
end

Ti?n extends
through the
thick laments

Nebulin binds
ac?n lament
and determines
the length
Tropomyosin and troponin are regulatory complexes associated with the thin lament

TM in its o state shiMs to its new posi?on (arrow) in the on state (right) when the
Ca2 concentra?on increases. This movement exposes myosin-binding sites (red) on
ac?n.
Regula?on of skeletal muscle contrac?on by Ca2 binding to TN. The TM-TN complex
remains bound to the thin lament whether muscle is relaxed or contracted.
Ac#n forms the contrac#le ring during cytokinesis
Yeasts use ac?n-myosin to
move organelles around,
microtubules more
common in animal cells
 Cytoplasmic streaming in giant algal cells: means of distribu#on of cellular metabolites

The center of a Nitella cell is lled with a single large water-lled vacuole, which is surrounded
by a layer of moving cytoplasm (indicated by blue arrows). A nonmoving layer of cor?cal
cytoplasm lled with chloroplasts lies just under the plasma membrane. On the inner side of
this layer are bundles of sta?onary ac?n laments (red), all oriented with the same polarity. A
motor protein (blue dots), most likely myosin XI, carries parts of the ER along the ac?n
laments. The movement of the ER network propels the en?re viscous cytoplasm, including
organelles that are enmeshed in the ER network.
Signal mediated rearrangement of ac#n laments
 SUMMARY

ac?n cytoskeleton is dynamic, with laments able to grow and shrink rapidly
Assembly, length, and stability of ac?n laments are controlled by specialized ac?n-
binding proteins that can sever laments or cap the ends or both.
These proteins are in turn regulated by various mechanisms.
All myosin isoforms can interact with ac?n laments through their head domains, but
their cellular roles dier, depending on their tail domains
Movement of ac?n laments by myosin can be directly monitored in the sliding-
lament assay
Myosins I, V, and VI power intracellular transloca?on of some membrane-limited
vesicles along ac?n laments.
A similar process is responsible for cytoplasmic streaming, which is probably mediated
by myosin XI, one of the fastest moving myosin
The contrac?le ring, a transient bundle of ac?n and myosin II, forms in a dividing cell
and pinches the cell into two halves in cytokinesis.
In skeletal muscle cells, ac?n thin laments and myosin thick laments are organized
into highly ordered structures called sarcomeres
force generated by myosin movement pulls the thin laments toward
the center of the sarcomere, shortening its length rapid rise in cytosolic Ca2 induced by
nerve s?mula?on of a skeletal muscle changes the interac?on between ac?n laments
and tropomyosin, exposing the myosin binding sites and thus permivng contrac?on to
occur