Cell Bio

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link the filaments to one another or to other cell
components such as plasma membrane, or influence the rate and extent of the filament polymerization.

Back 1

accessory proteins

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Primary antibodies bind specifically to cytoskeletal proteins.
Secondary antibodies labeled with a fluorescent tag bind to the primary ones, causing the cytoskeletal proteins to glow in the fluorescence microscope

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Immunofluorescence microscopy

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study in living cells: Fluorescent versions of cytoskeletal proteins are made and introduced into living cells. Fluorescence microscopy and video cameras are used to view the proteins as they function in the cells. GFP (green fluorescent protein)

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Fluorescence techniques

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High-resolution images from a video camera attached
to a microscope are computer processed to increase contrast and remove background features
that obscure the image. in vitro motility assay-motor protein activity; optical tweezers.
Confocal microscope

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Computer-enhanced video microscopy

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EM can resolve individual filaments prepared by thin section, quickfreeze
deep-etch, or negative staining techniques

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Electron microscopy

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Functions of a gene can be determined by studying the phenotype of the
cells/animals in which a particular gene product is either absent (knockout) or non-functional
(dominant-negative mutant).

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Genetic engineering

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?

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Biochemical characterization

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highly organized, stable MTs found in cilia, and flagella

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Axonemal MTs

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a network of MTs that extend from the perinuclear region of the cell in a
radial array.

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Cytoplasmic MTs

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Functions of cytoplasmic MTs

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determine the cell shape (axons)
-orientation of cellulose deposition during plant cell growth
-motile machinery

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Cytoskeleton General Functions

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supportive framework for
the cell, machinery for
coordinated movements, organize cell contents, transduce signals from extracellular
matrix and vise versus,anchoring
mRNAs

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Intermediate filaments

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Ropelike fibers,Eight
protofilaments joined end-to end with
staggered
overlaps, only non-polar
Ex. Keratin, Vimentin
Desmin

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Microtubules-

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Long,Hollow tube with a wall consisting of 13 protofilaments
Ex. α- and β-tubulin
Enzymatic Activity: GTPase
Motile machinery for organelle & chromosome movement, Maintenance of
axons

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Microfilaments

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Linear bundles,2D networks,3D actin gels. Two
intertwined,helical chains of actin
Ex. G-actin
Enzymatic Activity: ATPase

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Microtubule-associated proteins (MAPs):
MAP2C
Tau

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Tau: nerve axons; Bundles MTs; induces tubulin
polymerization
MAP2C:Nerve dendrites and cell body; Bundles MTs; increases MT stability and stiffness

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causes Alzheimer’s
disease

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Abnormal expression of
Tau is correlated with
neurodegenerative
disease

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MT assembly

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MTs are composed of subunits made of one molecule of
α-tubulin and one of β-tubulin : the tubulin heterodimer
dimers aggregate forming oligomers
Oligomers form linear chains: protofilaments
protofilaments form into sheets and 13+ close up into a tube (a MT)

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Critical concentration=?

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[tubulin] at steady-state.
nucleation->elongation->steady-state

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All anti-mitosis drugs which prevent cell division (like anti-cancer drugs)
MT depolymerizing agents:

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Colchicine (alkaloid from a plant), Colcemid
(colchicine derivative)
• Nocodazole
• Vincristine, Vinblastine
Taxol: from the yew. Stabilizes MT;
arrests cells at mitosis