BIO 201 Lecture Notes - Lecture 9: Gel Electrophoresis, Polyacrylamide, Microtubule
3 Apr 2017
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Cytoskeleton is a large structure, not very soluble: microtubules, microfilaments, intermediate filaments. Characterization of cytoskeletal proteins using sds polyacrylamide gel electrophoresis (sds-page: homogenize cells, purify protein, denature protein- have to add sds. Sds (detergent; disrupts non covalent interactions; binds to proteins giving them negative charge) Heat (works with sds to disrupt non covalent interactions: sds page. Sds-page: protein, add sds + bme and boil, large proteins migrate slowly, small proteins migrate faster, anode. Three types of microtubules: cytoplasmic-cytoplasm, unstable, axonemal-cilia and flagella, stable, centriolar-centrioles, stable. A and b tubulin subunits form heterodimers. Polymers only in the presence of gtp. Mts are polar: plus (+) end: polymerization favorable minus end: unfavorable. *tubulin catalyzes hydrolysis of gtp; it is a gtpase. Mts are in dynamic equilibrium of polymerization and depolymerization at all times. Made from centriole (mts) and other proteins. Is microtubule organizing center in plants and animals.
