Differential interaction of tubulin isotypes with the antimitotic compound IKP-104.
The tubulin molecule is a heterodimer composed of two polypeptide chains, designated alpha and beta; both alpha and beta exist in numerous isotypic forms, which differ in their assembly and drug binding properties. 2-(4-Fluorophenyl)-1-(2-chloro-3, 5-dimethoxyphenyl)-3-methyl-6-phenyl-4(1H)-pyridinone (IKP-104) is an antimitotic compound which inhibits polymerization and induces depolymerization of microtubules [Mizuhashi, F., et al. (1992) Jpn. J. Cancer Res. 83, 211]. Since the previous work was undertaken with isotypically unfractionated tubulin, we have investigated the interactions of IKP-104 with the isotypically purified tubulin dimers (alpha beta(II), alpha beta(III), and alpha beta(IV)). We find that IKP-104 binds to alpha beta(II) and alpha beta(III) at two classes of binding sites. However, affinities for each class of site are much weaker for alpha beta(III) than for alpha beta(II). Interestingly, the low-affinity site on alpha beta(IV) was not detectable. Its high-affinity site was weaker than those of either alpha beta(II) or alpha beta(III). In a pattern consistent with these results, IKP-104 inhibited assembly better with alpha beta(II) than with the other two dimers. Higher concentrations of IKP-104 induced formation of spiral aggregates from alpha beta(II) and alpha beta(III) but not from alpha beta(IV). Our results suggest that the interaction of IKP-104 with tubulin isotypes is very complex: alpha beta(II) and alpha beta(III) differ quantitatively in their interaction with IKP-104, and alpha beta(IV)'s interaction differs both quantitatively and qualitatively from those of the other two dimers.
Department of Biochemistry, University of Texas Health Science Center, San Antonio, Texas 78284, USA. israr@bioc02.UTHSCSA.edu
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Khan
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Tomita
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Mizuhashi
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Ludueña
D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism / Q000494:pharmacology*; D002417:Cattle; D019281:Dimerization; D004305:Dose-Response Relationship, Drug; D007700:Kinetics; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism; D020033:Protein Isoforms; D011728:Pyridones / Q000378:metabolism / Q000494:pharmacology*; D013050:Spectrometry, Fluorescence; D014404:Tubulin / Q000378:metabolism*
D000970:Antineoplastic Agents; D020033:Protein Isoforms; D011728:Pyridones; D014404:Tubulin; C067096:IKP 104