Mass spectrometry in coupling with affinity capture-release and isotope-coded affinity tags for quantitative protein analysis

被引:58
作者
Turecek, F [1 ]
机构
[1] Univ Washington, Dept Chem, Seattle, WA 98195 USA
来源
JOURNAL OF MASS SPECTROMETRY | 2002年 / 37卷 / 01期
关键词
affinity capture; biotinylated conjugates; isotope labeling; electrospray ionization; genetic disorders; proteomics;
D O I
10.1002/jms.275
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Affinity capture-release electrospray ionization mass spectrometry (ACESIMS) and isotope-coded affinity tags (ICAT) are two recently introduced techniques for the quantitation of protein activity and content with applications to clinical enzymology and functional proteomics, respectively. One common feature of these methods is that they use biotinylated tags that function as molecular handles for highly selective and reversible affinity capture of conjugates from complex biological mixtures such as cell homogenates and sub-cellular organelles. ACESIMS uses synthetic substrate conjugates specifically to target cellular enzymes that, when deficient, are the cause of genetic diseases. Multiplex determination of enzyme activities is used for the diagnosis of lysosomal storage diseases. The ICAT method relies on selective conjugation of cysteine thiol groups in proteins, followed by enzymatic digestion and quantitative analysis of peptide conjugates by mass spectrometry. Another common feature of the ACESIMS and ICAT approaches is that both use conjugates labeled with stable heavy isotopes as internal standards for quantitation. Selected applications of the ACESIMS and ICAT techniques are presented that include molecular-level diagnosis of genetic diseases in children and quantitative determination of protein expression in cells. Copyright (C) 2001 John Wiley Sons, Ltd.
引用
收藏
页码:1 / 14
页数:14
相关论文
共 77 条
[1]   Selective analysis of phosphopeptides within a protein mixture by chemical modification, reversible biotinylation and mass spectrometry [J].
Adamczyk, M ;
Gebler, JC ;
Wu, J .
RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 2001, 15 (16) :1481-1488
[2]  
[Anonymous], METHODS ENZYMOLOGY, DOI [10.1016/0076-6879(87)48036-1, DOI 10.1016/0076-6879(87)48036-1]
[3]  
ARUR S, 2001, P 49 ASMS C MASS SPE
[4]  
BURKHART W, 2001, P 49 ASMS C MASS SPE
[5]   Inhibition of glycosphingolipid biosynthesis: Application to lysosomal storage disorders [J].
Butters, TD ;
Dwek, RA ;
Platt, FM .
CHEMICAL REVIEWS, 2000, 100 (12) :4683-+
[6]  
CHACE DH, 1993, CLIN CHEM, V39, P66
[7]  
Chen Y.T., 1995, METABOLIC MOL BASES, P935
[8]  
CONRADS TP, 2001, P 49 ASMS C MASS SPE
[9]  
Corthals GL, 2000, ELECTROPHORESIS, V21, P1104, DOI 10.1002/(SICI)1522-2683(20000401)21:6<1104::AID-ELPS1104>3.0.CO
[10]  
2-C