Possible regulation of the in vitro assembly of bovine brain tubulin by the bovine thioredoxin system.
Microtubule assembly in vitro and in vivo is highly sensitive to a variety of sulfhydryl-reactive reagents, raising the question of the possible existence of a physiological sulfhydryl-mediated system for regulating microtubule assembly. However, the specific reagents which have previously been used to inhibit microtubule assembly in vitro are either nonphysiological or, if physiological, effective only at concentrations much higher than their physiological ones. Because of reports of association in vivo between microtubules and the sulfhydryl-reactive proteins thioredoxin and thioredoxin reductase, we decided to examine the interaction in vitro between microtubules and the thioredoxin system, comprising thioredoxin, thioredoxin reductase and NADPH. At pH 6.8, both the mammalian and the Escherichia coli thioredoxin systems inhibited microtubule assembly by 4-35% (19 +/- 9%) by reducing one intra-subunit disulfide bond in the tubulin dimer. The thioredoxin-reducible disulfide of the tubulin dimer remains protected from thioredoxin in the assembled microtubules. Thioredoxin or thioredoxin reductase alone, or together in the absence of NADPH, were incapable of either reducing tubulin or inhibiting microtubule assembly. Microtubules formed from reduced tubulin were found to be stable and morphologically identical to those obtained from native tubulin dimers. Since the components of the thioredoxin system were used at concentrations similar to their physiological ones, our results suggest a potential role of the thioredoxin system in regulation of microtubule assembly in vivo.
Khan
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IA
Department of Biochemistry, University of Texas Health Science Center, San Antonio 78284-7760.
Ludueña
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RF
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008870:Microtubules / Q000378:metabolism* / Q000648:ultrastructure; D013880:Thioredoxin-Disulfide Reductase / Q000302:isolation & purification / Q000378:metabolism; D013879:Thioredoxins / Q000302:isolation & purification / Q000378:metabolism*; D014404:Tubulin / Q000378:metabolism* / Q000648:ultrastructure
D046911:Macromolecular Substances; D014404:Tubulin; D013879:Thioredoxins; D013880:Thioredoxin-Disulfide Reductase