title: Methods for studying microtubule binding site interactions: zampanolide as a covalent binding agent.
abstract: 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.
authors: School of Biological Sciences, Victoria University of Wellington, Wellington, New Zealand.
authors: Jessica J
authors: JJ
authors: Field
authors: Enrique
authors: E
authors: Calvo
authors: Peter T
authors: PT
authors: Northcote
authors: John H
authors: JH
authors: Miller
authors: Karl-Heinz
authors: KH
authors: Altmann
authors: José Fernando
authors: JF
authors: Díaz
mesh_terms: D000818:Animals; D001665:Binding Sites / Q000502:physiology*; D001921:Brain / Q000166:cytology / Q000378:metabolism; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000378:metabolism; D002417:Cattle; D007783:Lactones / Q000378:metabolism; D018942:Macrolides / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D017239:Paclitaxel / Q000378:metabolism; D011083:Polycyclic Compounds / Q000378:metabolism; D011485:Protein Binding / Q000502:physiology*; D050257:Tubulin Modulators / Q000378:metabolism
chemical_list: D019086:Bridged Bicyclo Compounds, Heterocyclic; C408303:FR 182877; D007783:Lactones; D018942:Macrolides; D011083:Polycyclic Compounds; D050257:Tubulin Modulators; C117305:laulimalide; C414085:peloruside A; C444900:zampanolide; D017239:Paclitaxel
keywords: Anticancer drugs; Covalent binding drug; Flutax; Microtubule-stabilizing agent; Taxoid site; Zampanolide