Structure and importance of the dimerization domain in elongation factor Ts from Thermus thermophilus

被引:38
作者
Jiang, YX
Nock, S
Nesper, M
Sprinzl, M
Sigler, PB
机构
[1] YALE UNIV, DEPT MOL BIOPHYS & BIOCHEM, NEW HAVEN, CT 06520 USA
[2] YALE UNIV, DEPT CHEM, NEW HAVEN, CT 06520 USA
[3] YALE UNIV, HOWARD HUGHES MED INST, NEW HAVEN, CT 06520 USA
[4] UNIV BAYREUTH, BIOCHEM LAB, D-8580 BAYREUTH, GERMANY
关键词
D O I
10.1021/bi960918w
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
Elongation factor Ts (EF-Ts) functions as a nucleotide-exchange factor by binding elongation factor Tu (EF-Tu) and accelerating the GDP dissociation from EF-Tu; thus EF-Ts promotes the transition of EF-Tu from the inactive GDP form to the active GTP form. Thermus thermophilus EF-Ts exists as a stable dimer in solution which binds two molecules of EF-Tu to form a (EF-Tu . EF-Ts)(2) heterotetramer. Here we report the crystal structure of the dimerization domain of EF-Ts from T. thermophilus refined to 1.7 Angstrom resolution. A three-stranded antiparallel beta-sheet from each subunit interacts to form a beta-sandwich that serves as an extensive dimer interface tethered by a disulfide bond. This interface is distinctly different from the predominantly alpha-helical one that stabilizes the EF-Ts dimer from Escherichia coli [Kawashima, T., et al. (1996) Nature 379, 511-518]. To test whether the homodimeric form of T. thermophilus EF-Ts is necessary for catalyzing nucleotide exchange, the present structure was used to design mutational changes within the dimer interface that disrupt the T. thermophilus EF-Ts dimer but not the tertiary structure of the subunits. Surprisingly, EF-Ts monomers created in this manner failed to catalyze nucleotide exchange in EF-Tu, indicating that, in vitro, T. thermophilus EF-Ts functions only as a homodimer.
引用
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页码:10269 / 10278
页数:10
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