Tubulin subunit carboxyl termini determine polymerization efficiency

Abstract

Cleavage of tubulin by subtilisin removes a small (M_r < 2000) fragment from the C-terminal end of both α and β subunits. The resulting protein is much reduced in negative charge. The cleaved, less acidic protein retains its competence to polymerize in a GTP-dependent and cold-, GDP-, and podophyllotoxin-sensitive manner and assembles into sheets or bundles of twisted filaments. The critical concentration for polymerization of the cleaved protein is about 50-fold lower than that for intact tubulin. It is proposed that the C termini of the subunits normally impede polymerization.