A generic protein purification method for protein complex characterization and proteome exploration

被引:2102
作者
Rigaut, G [1 ]
Shevchenko, A [1 ]
Rutz, B [1 ]
Wilm, M [1 ]
Mann, M [1 ]
Séraphin, B [1 ]
机构
[1] European Mol Biol Lab, D-69117 Heidelberg, Germany
关键词
D O I
10.1038/13732
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
We have developed a generic procedure to purify proteins expressed at their natural level under native conditions using a novel tandem affinity purification (TAP) tag. The TAP tag allows the rapid purification of complexes from a relatively small number of cells without prior knowledge of the complex composition, activity, or function. Combined with mass spectrometry, the TAP strategy allows for the identification of proteins interacting with a given target protein. The TAP method has been tested in yeast but should be applicable to other cells or organisms.
引用
收藏
页码:1030 / 1032
页数:3
相关论文
共 16 条
[1]   Partial purification of the yeast U2 snRNP reveals a novel yeast pre-mRNA splicing factor required for pre-spliceosome assembly [J].
Caspary, F ;
Shevchenko, A ;
Wilm, M ;
Séraphin, B .
EMBO JOURNAL, 1999, 18 (12) :3463-3474
[2]   MOLECULAR GENETIC-ANALYSIS OF A PLANT-VIRUS POLYPROTEIN CLEAVAGE SITE - A MODEL [J].
DOUGHERTY, WG ;
CARY, SM ;
PARKS, TD .
VIROLOGY, 1989, 171 (02) :356-364
[3]  
Ford C F, 1991, Protein Expr Purif, V2, P95, DOI 10.1016/1046-5928(91)90057-P
[4]  
FORTES P, 1999, IN PRESS GENES DEV, V3
[5]   Toward a functional analysis of the yeast genome through exhaustive two-hybrid screens [J].
FromontRacine, M ;
Rain, JC ;
Legrain, P .
NATURE GENETICS, 1997, 16 (03) :277-282
[6]   Importin provides a link between nuclear protein and U snRNA export [J].
Gorlich, D ;
Kraft, R ;
Kostka, S ;
Vogel, F ;
Hartmann, E ;
Laskey, RA ;
Mattaj, IW ;
Izaurralde, E .
CELL, 1996, 87 (01) :21-32
[7]  
Gottschalk A, 1998, RNA, V4, P374
[8]  
Puig O, 1998, YEAST, V14, P1139
[9]   SMALL NUCLEAR RNAS FROM SACCHAROMYCES-CEREVISIAE - UNEXPECTED DIVERSITY IN ABUNDANCE, SIZE, AND MOLECULAR COMPLEXITY [J].
RIEDEL, N ;
WISE, JA ;
SWERDLOW, H ;
MAK, A ;
GUTHRIE, C .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1986, 83 (21) :8097-8101
[10]   Mtr10p functions as a nuclear import receptor for the mRNA-binding protein Np13p [J].
Senger, B ;
Simos, G ;
Bischoff, FR ;
Podtelejnikov, A ;
Mann, M ;
Hurt, E .
EMBO JOURNAL, 1998, 17 (08) :2196-2207