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. EM 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 2 actins show a high degree of homology although apparently 5 of the 25 spots visualized by fluorescamine show a difference in chromatographic mobility. The purification procedure described allows the rapid isolation of actin and tubulin from tissue culture cells in sufficient amounts for comparative biochemical studies.