title: The isolation of tubulin and actin from mouse 3T3 cells transformed by simian virus 40 (SV3T3 cells), an established cell line growing in culture.
abstract: Tubulin can be purified from mouse SV3T3 cells (3T3 cells transformed by SV40 virus) by several cycles of temperature-dependent polymerization and depolymerization. Electron microscopical analysis of the final product reveals morphologically normal microtubules. Homogeneous actin can be isolated as a byproduct of the purification procedure. Mouse SV3T3 actin and skeletal muscle actin were compared by fingerprint analysis of the tryptic peptides obtained from performic-acid-oxidized protein. The two actins show a high degree of homology although apparently five of the twenty-five spots visualized by fluorescamine show a difference in chromatographic mobility. The purification procedure described allows the rapid isolation of both actin and tubulin from tissue culture in sufficient amounts for comparative biochemicals studies.
mesh_terms: D000199:Actins / Q000302:isolation & purification*; D002460:Cell Line; D002472:Cell Transformation, Viral; D006023:Glycoproteins / Q000302:isolation & purification*; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008870:Microtubules / Q000648:ultrastructure; D010446:Peptide Fragments / Q000032:analysis; D013539:Simian virus 40; D013696:Temperature; D014404:Tubulin / Q000302:isolation & purification*
chemical_list: D000199:Actins; D006023:Glycoproteins; D046911:Macromolecular Substances; D010446:Peptide Fragments; D014404:Tubulin