Volker K W, Knull H R
Department of Biochemistry and Molecular Biology, School of Medicine, University of North Dakota, Grand Forks 58202.
J Mol Recognit. 1993 Dec;6(4):167-77. doi: 10.1002/jmr.300060405.
Tubulin and microtubules were modified with the protease, subtilisin. The modification reduced the length of alpha- or beta-tubulin by cleaving a peptide fragment from the C-terminals. Generation of alpha'beta'-tubulin, which is cleaved at both the alpha- and beta-subunit terminals, and alpha beta'-tubulin, which is cleaved at the beta-subunit C-terminal, have already been reported. In this work an isotype, alpha'beta-tubulin, was produced. The three modified tubulin isotypes were compared for their ability to interact with glycolytic enzymes. Cleavage of alpha led to a poorer interaction when tested via affinity chromatography. Tubulin also inhibits the activity of aldolase and glyceraldehyde 3-phosphate dehydrogenase. When the alpha-subunit C-terminal was intact, inhibition was greatest. These results imply that the C-terminal of the tubulin alpha-subunit is responsible for interactions with glycolytic enzymes.
微管蛋白和微管被蛋白酶枯草杆菌蛋白酶修饰。这种修饰通过从C末端切割肽片段来缩短α-或β-微管蛋白的长度。已经报道了在α-和β-亚基末端均被切割的α'β'-微管蛋白以及在β-亚基C末端被切割的αβ'-微管蛋白的产生。在这项工作中，产生了一种同种型，即α'β-微管蛋白。比较了这三种修饰的微管蛋白同种型与糖酵解酶相互作用的能力。通过亲和色谱法测试时，α的切割导致相互作用较差。微管蛋白还抑制醛缩酶和3-磷酸甘油醛脱氢酶的活性。当α-亚基C末端完整时，抑制作用最大。这些结果表明，微管蛋白α-亚基的C末端负责与糖酵解酶的相互作用。
