Lecture 6: Cytoskeleton Flashcards
Cytoskeleton is made up of these three specific protein structures
Microfilaments (actin)-7nm thick
Intermediate filaments- 8-10nm thick
Microtubules- 25nm in diameter
Characteristics of Actin
Highly conserved among eukaryotes
7nm thick, up to 7um in length
Organized into bundles and 3D networks
Bind to specific transmembrane proteins directly or indirectly
Exist as monomers (G-actin) and long chains (F-actin)
Why do actin filaments display polarity
Because all actin monomers are oriented in the same direction
First step in actin polymerization
Nucleation: a trimer is formed, then additional actin can be added to either side
The growing end is barbed or pointy
Barbed
Treadmilling
Intermediate concentrations of G-actin favor a dynamic equilibrium between the +/- end. No net growth
Low concentrations of G-actin favor
Depolymerization of actin filaments
Higher concentrations of G-actin favor
Net addition of actin at both ends, therefore growing the actin filament
Cytochalasins
Bind to barbed ends of actin, block elongation
Can inhibit movements (e.g., cell division)
Phalloidin
Binds to actin filaments and prevents dissociation
Can be labeled with fluorescent dyes to allow visualization of actin filaments
Diversity of function of actin filaments in a cell is determined by
Actin-binding proteins
Spectrin
Found in RBCs
Binds cortical cytoskeleton to plasma membrane
Dystrophin
Binds cortical cytoskeleton to plasma membrane
Villin and Fimbrin
Cross-links actin in microvilli
Calmodulin and Myosin I
Cross-links actin to plasma membrane in microvilli
alpha-Actinin
Cross-links stress fibers and connects actin to protein-plasma membrane complexes
Filamin
Cross-links actin at wide angels to form screen-like gels
Thymosin
Captures/bind actin monomers, prevents them from being polymerized
Profilin
Binds to actin monomers and prevents them from being polymerized
Facilitates exchange of bound ADP for ATP-which favor polymerization
Gelsolin
Destabilizes F-actin and caps actin filaments, preventing loss and addition of G-actin
In presence of calcium ion, it fragments actin filaments and stays bound to +end
Cofilin
Triggers depolymerization of ADP-bound actin at the minus end
Arp2/3
Initiates polymerization of F-actin from sides of existing filament (causes branching)
Latrunculins
Binds to G-actin and induces F-actin depolymerization
How thick are intermediate filaments
8-10nm
Intermediate filaments are abundant in cells subjected to
Mechanical stress
Intermediate filaments strengthen epithelial cells as
Desmosomes and hemidesmosomes
Intermediate filament central unit
Alpha-helical rod flanked by head and tail domains
Head and tail domains of intermediate filaments determine
Specific functions
Intermediate filament functions
Form a cytoplasmic network in most cells
Associate with other cytoskeletal elements to form a scaffolding that organizes the internal structure of a cell
Dimer
Central rod of two polypeptides together
Rods are alined tail-tail, head-head
Tetramer
Dimers associated in a staggered anti-parallel fashion
Why do intermediate filaments not exhibit behaviors such as treadmilling
They are more stable than actin because they do not have distinct ends, due to the antiparallel association of dimers
Protofilament
Tetramers assemble end to end
Filament
Eight protofilaments wound together
Most common type of intermediate filament
Keratin
Microtubule thickness
25nm
Microtubules are composed of what units
alpha and beta units
Protofilaments are
longitudinal rows of tubulin dimers
How many protofilaments arrange in what orientation for microtubules
13 parallel protofilaments
____ dimers with ___ bound to the _____ associate with the growing end
Tubulin dimers with GTP bound to the beta unit
Plus end grows more rapidly than minus end in presence of
low calcium ion concentration
After polymerization, ___ is hydrolyzed to ___ and the tubulin is ____ stable
GTP is hydrolyzed to GTP and tubulin is less stable
Dynamic instability
Individual microtubules undergo alternate phases of slow growth and rapid depolymerization
alpha tubulin associates with the ___ end, beta tubulin associates with the ___ end
alpha tubulin=minus end
beta tubulin=plus end
Colchicines
Inhibit microtubule polymerization
Colcemid
inhibit microtubule polymerization
Vincristine
inhibit microtubule polymerization
Taxol
Stabilize microtubules
Vinblastine
Inhibit microtubule polymerization
Nocodazole
Inhibits microtubule polymerization
Cytoskeleton functions (7)
Cell movement Support and strength for cell Phagocytosis Mitotic spindle formation Cytokenisis Cell-cell, cell-extracellular matrix adherence Changes in cell shape
Kinesin
Anterograde transport protein
Dynein
Retrograde transport protein
Raft protein complex
May provide a mechanism for transport of multiple cargos
Myosin I number of heads, tail binds to what, head binds to what, direction of head motion towards the
One head
Tail binds to cell membrane
Head binds to actin
Head motion towards barbed end
Myosin II number of heads, tail binds to what, head binds to what, direction of head motion towards the
Two heads
Tail binds to Myosin II
Head binds to Actin
Head motion towards barbed end