The isolation of tubulin and actin from mouse 3T3 cells transformed by simian virus 40 (SV3T3 cells), an established cell line growing in culture.
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.
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*
D000199:Actins; D006023:Glycoproteins; D046911:Macromolecular Substances; D010446:Peptide Fragments; D014404:Tubulin