鉴定微管蛋白上的12Cysbeta作为微管嗪的结合位点。
Identification of 12Cysbeta on tubulin as the binding site of tubulyzine.
作者信息
Kim Yeoun Jin, Sackett Dan L, Schapira Matthieu, Walsh Daniel P, Min Jaeki, Pannell Lewis K, Chang Young-Tae
机构信息
National Institute of Diabetes, Digestive, Kidney Diseases, National Institutes of Health, US Department of Health and Human Services, Bethesda, MD 20892, USA.
出版信息
Bioorg Med Chem. 2006 Feb 15;14(4):1169-75. doi: 10.1016/j.bmc.2005.09.069. Epub 2005 Nov 2.
We have undertaken quantitative binding site studies in order to identify the binding site of the known microtubule destabilizing agents, the tubulyzines, in the tubulin dimer. Two different approaches were employed that utilized the tubulyzines and their derivatives. The first approach was based on a chemical affinity labeling method using tubulyzine affinity derivatives, and the second approach employed the mass spectrometric measurement of the differential reactivity of cysteines using the tubulyzines and monobromobimane. Based on overlapping data from these two approaches, we propose that the tubulyzines bind at the guanosine-5'-triphosphate binding site of beta-tubulin. Interestingly, we also show that the tubulyzines' binding to tubulin induces a conformational change in tubulin that prevents further interaction of the 239Cysbeta with other reagents.
为了确定已知的微管解聚剂tubulyzines在微管蛋白二聚体中的结合位点，我们进行了定量结合位点研究。采用了两种不同的方法，它们都利用了tubulyzines及其衍生物。第一种方法基于使用tubulyzine亲和衍生物的化学亲和标记法，第二种方法则利用tubulyzines和单溴代双马来酰亚胺通过质谱测量半胱氨酸的差异反应性。基于这两种方法的重叠数据，我们提出tubulyzines结合在β-微管蛋白的鸟苷-5'-三磷酸结合位点。有趣的是，我们还表明tubulyzines与微管蛋白的结合会诱导微管蛋白发生构象变化，从而阻止239Cysβ与其他试剂的进一步相互作用。
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