A novel multiple affinity purification tag and its use in identification of proteins associated with a cyclin-CDK complex

被引：68

作者：

Honey, Sangeet ^{[1
]}

Schneider, Brandt L. ^{[1
]}

Schieltz, David M. ^{[2
]}

Yates, John R. ^{[2
]}

Futcher, Bruce ^{[1
]}

机构：

[1] Cold Spring Harbor Labs, Cold Spring Harbor, NY 11724 USA

[2] Univ Washington, Dept Mol Biotechnol, Hlth Sci Ctr, Seattle, WA 98195 USA

来源：

NUCLEIC ACIDS RESEARCH | 2001年 / 29卷 / 04期

基金：

美国国家卫生研究院;

关键词：

D O I：

10.1093/nar/29.4.e24

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：

A novel multiple affinity purification (MAFT) or tandem affinity purification (TAP) tag has been constructed. It consists of the calmodulin binding peptide, six histidine residues, and three copies of the hemagglutinin epitope. This 'CHH' MAFT tag allows two or three consecutive purification steps, giving high purity. Active Clb2-Cdc28 kinase complex was purified from yeast cells after inserting the CHH tag into Clb2. Associated proteins were identified using mass spectrometry. These included the known associated proteins Cdc28, Sic1 and Cks1. Several other proteins were found including the 70 kDa chaperone, Ssa1.

引用

页数：9

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