Field Jessica J, Calvo Enrique, Northcote Peter T, Miller John H, Altmann Karl-Heinz, Díaz José Fernando
School of Biological Sciences, Victoria University of Wellington, Wellington, New Zealand.
Methods Cell Biol. 2013;115:303-25. doi: 10.1016/B978-0-12-407757-7.00019-0.
In this chapter, we describe the methods used to determine the binding site and binding profile of zampanolide, a novel microtubule-stabilizing agent (MSA) that binds covalently to tubulin. These methods can be applied to other novel MSAs in which the binding site and mechanism of binding are unknown. Using the described methods, we have shown that zampanolide binds to the taxoid site on β-tubulin, but unlike most other MSAs is able to covalently modify this site. The purpose of this chapter is to provide a step-by-step protocol for determining the binding site of a novel MSA.
在本章中，我们描述了用于确定扎姆帕诺利德（一种与微管蛋白共价结合的新型微管稳定剂）结合位点和结合模式的方法。这些方法可应用于其他结合位点和结合机制未知的新型微管稳定剂。使用所述方法，我们已表明扎姆帕诺利德与β-微管蛋白上的紫杉烷类位点结合，但与大多数其他微管稳定剂不同的是，它能够共价修饰该位点。本章的目的是提供一个用于确定新型微管稳定剂结合位点的详细步骤方案。
