The yeast Ski complex is a hetero-tetramer

被引：39

作者：

Synowsky, Silvia A. ^{[1
,2
]}

Heck, Albert J. R. ^{[1
,2
]}

机构：

[1] Univ Utrecht, Bijvoet Ctr, Biomol Res & Utrecht Inst Pharmaceut Sci, Dept Biomol Mass Spectrometry, NL-3584 CA Utrecht, Netherlands

[2] Univ Utrecht, Utrecht Inst Pharmaceut Sci, NL-3584 CA Utrecht, Netherlands

来源：

PROTEIN SCIENCE | 2008年 / 17卷 / 01期

关键词：

Ski complex; exosome; native mass spectrometry; protein complex structure and topology;

D O I：

10.1110/ps.073155908

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 yeast Ski complex assists the exosome in the degradation of mRNA. The Ski complex consists of three components; Ski2, Ski3, and Ski8, believed to be present in a 1:1:1 stoichiometry. Measuring the mass of intact isolated endogenously expressed Ski complexes by native mass spectrometry we unambiguously demonstrate that the Ski complex has a hetero- tetrameric stoichiometry consisting of one copy of Ski2 and Ski3 and two copies of Ski8. To validate the stoichiometry of the Ski complex, we performed tandem mass spectrometry. In these experiments one Ski8 subunit was ejected concomitant with the formation of a Ski2/Ski3/Ski8 fragment, confirming the proposed stoichiometry. To probe the topology of the Ski complex we disrupted the complex and mass analyzed the thus formed subcomplexes, detecting Ski8-Ski8, Ski2-Ski3, Ski8-Ski2, and Ski8-Ski8-Ski2. Combining all data we construct an improved structural model of the Ski complex.

引用

页码：119 / 125

页数：7

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