The cross-linking of tubulin with imidoesters.
Tubulin was reacted with a monofunctional imidoester, ethyl acetimidate, and three bifunctional imidoesters ranging in extension from 5 to 10 A. Extensive cross-linking was found to occur with the three bifunctional reagents resulting in the formation of dimers, trimers, tetramers, pentamers, and hexamers. In addition, a similar cross-linking patterns was observed with the monofunctional reagent. This type of cross-linking is rarely seen when proteins are reacted with imidoesters. Given that the cross-linking span of ethyl acetimidate is only 3 A it is reasonable to infer that there are nucleophilic groups in close proximity within the tubulin dimer. Amidinated tubulin is still capable of assembling into microtubules.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007096:Imidoesters; D008870:Microtubules / Q000032:analysis / Q000648:ultrastructure; D009391:Nephelometry and Turbidimetry / Q000379:methods; D013552:Swine; D014404:Tubulin; D014461:Ultracentrifugation
D000596:Amino Acids; D003432:Cross-Linking Reagents; D007096:Imidoesters; D014404:Tubulin