DOMAIN-STRUCTURE OF ESCHERICHIA-COLI DNA GYRASE AS REVEALED BY DIFFERENTIAL SCANNING CALORIMETRY

被引:18
作者
BLANDAMER, MJ
BRIGGS, B
CULLIS, PM
JACKSON, AP
MAXWELL, A
REECE, RJ
机构
[1] UNIV LEICESTER, DEPT CHEM, LEICESTER LE1 7RH, ENGLAND
[2] UNIV LEICESTER, DEPT BIOCHEM, LEICESTER LE1 7RH, ENGLAND
基金
英国惠康基金;
关键词
D O I
10.1021/bi00190a003
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The domain structure of DNA gyrase from Escherichia coli has been examined using differential scanning microcalorimetry. The intact enzyme (an A(2)B(2) tetramer) shows at least four transitions with apparent T-m's at 44.8, 53.3, 58.6, and 60.7 degrees C. Comparison with the thermal stabilities of the two separate subunits and genetically-engineered protein fragments has been used to assign these transitions to individual domains within the intact gyrase proteins. The thermal unfolding of DNA gyrase and all individual fragments are irreversible under the conditions of the calorimetric experiment. Further evidence for the assignment of transitions to particular domains has been obtained by studying the effects of tight-binding ligands such as novobiocin on the thermal stabilities of the various protein fragments.
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页码:7510 / 7516
页数:7
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