Cryptophycin 1 binds to tubulin at a site distinct from the colchicine binding site and at a site that may overlap the vinca binding site.

Cryptophycin 1 is a new cytotoxic antimicrotubule agent with excellent antitumor activity. The methods of Sackett (Biochemistry, 34, 7010-7019, 1995), utilizing the selective and specific proteolysis of alpha- and beta-tubulin by trypsin and chymotrypsin, was used to identify the cryptophycin 1 binding site on tubulin. Occupancy of the colchicine or vinca binding sites causes changes in the structure of tubulin that can be detected by proteolysis with trypsin and chymotrypsin. The addition of cryptophycin 1 to tubulin causes changes in both the tryptic and chymotryptic cleavage of tubulin consistent with occupation of the vinca binding site and distinct from occupation of the colchicine binding site. The effects of cryptophycin 1 on the tryptic digests are identical to the effects seen with vinblastine and differ saliently from the effects of maytansine and rhizoxin, other agents known to bind to the vinca site. The data suggest that the binding site of cryptophycin 1 may overlap the vinca binding site on tubulin.