Localization of the exchangeable nucleotide binding domain in beta-tubulin.
Limited proteolysis of tubulin by alpha-chymotrypsin cleaved the beta-subunit preferentially at Tyr 281, generating primarily 35 kD and 17 kD fragments which were located in the amino terminal and the carboxy terminal regions, respectively. A small amount of a 19 kD fragment from the C-terminal end was also produced. Alpha-Chymotrypsin-treated tubulin retained the ability to exchange GTP and covalently incorporate nucleotide by direct photoaffinity labeling. SDS-PAGE and autoradiography analysis of the [alpha-32P] GTP-labeled alpha-CT-treated tubulin showed that the 35 kD fragment was almost exclusively labeled, indicating that the exchangeable GTP binding domain resides in the amino terminal region of the beta-subunit.
D001665:Binding Sites; D002918:Chymotrypsin / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D011108:Polymers; D014404:Tubulin / Q000378:metabolism*; D014466:Ultraviolet Rays
D010446:Peptide Fragments; D011108:Polymers; D014404:Tubulin; D006160:Guanosine Triphosphate; D002918:Chymotrypsin