Efficient enrichment of intact phosphorylated proteins by modified immobilizes metal-affinity chromatography

被引：53

作者：

Dubrovska, A ^{[1
]}

Souchelnytskyi, S ^{[1
]}

机构：

[1] Uppsala Univ, Ctr Biomed, Ludwig Inst Canc Res, SE-75124 Uppsala, Sweden

来源：

PROTEOMICS | 2005年 / 5卷 / 18期

关键词：

immobilized metal-affinity chromatography; phosphoproteomics; phosphorylation; two-dimensional gel electrophoresis;

D O I：

10.1002/pmic.200500002

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Phosphoproteome studies are hampered by the lack of methods which allow a comprehensive Received: January 4, 2005 and fast analysis of intact phosphoproteins. Here we describe an immobilized metal-affinity Revised: April 16, 2005 chromatography (IMAC)-based technique for the enrichment of phosphorylated proteins, which Accepted: April 25, 2005 allows recovery of up to 90% of phosphoproteins. This technique is compatible with 2-DE and can be applied to cultured cells and tissues.

引用

页码：4678 / 4683

页数：6

共 21 条

[1] Chemical strategies for functional proteomics [J].

Adam, GC ;

Sorensen, EJ ;

Cravatt, BF .

MOLECULAR & CELLULAR PROTEOMICS, 2002, 1 (10) :781-790

[2] Actin cytoskeletal association of cytohesin-1 is regulated by specific phosphorylation of its carboxyl-terminal polybasic domain [J].

Dierks, H ;

Kolanus, J ;

Kolanus, W .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (40) :37472-37481

[3] Phosphoproteome analysis by mass spectrometry and its application to Saccharomyces cerevisiae [J].

Ficarro, SB ;

McCleland, ML ;

Stukenberg, PT ;

Burke, DJ ;

Ross, MM ;

Shabanowitz, J ;

Hunt, DF ;

White, FM .

NATURE BIOTECHNOLOGY, 2002, 20 (03) :301-305

[4]

Figeys D, 2002, ANAL CHEM, V74, p413A

[5]

Imam-Sghiouar N, 2002, PROTEOMICS, V2, P828, DOI 10.1002/1615-9861(200207)2:7<828::AID-PROT828>3.0.CO

[6]

2-T

[7] Functional proteomics of transforming growth Factor-β1-stimulated Mv1Lu epithelial cells:: Rad51 as a target of TGFβ1-dependent regulation of DNA repair [J].

Kanamoto, T ;

Hellman, U ;

Heldin, CH ;

Souchelnytskyi, S .

EMBO JOURNAL, 2002, 21 (05) :1219-1230

[8] Phosphospecific proteolysis for mapping sites of protein phosphorylation [J].

Knight, ZA ;

Schilling, B ;

Row, RH ;

Kenski, DM ;

Gibson, BW ;

Shokat, KM .

NATURE BIOTECHNOLOGY, 2003, 21 (09) :1047-1054

[9] Agonist-induced tyrosine phosphorylation of Gq/G11α requires the intact structure of membrane domains [J].

Matousek, P ;

Durchánková, D ;

Svandová, I ;

Novotny, J ;

Svoboda, P .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2005, 328 (02) :526-532

[10] Cell-cycle dependent tyrosine phosphorylation on mortalin regulates its interaction with fibroblast growth factor-1 [J].

Mizukoshi, E ;

Suzuki, M ;

Misono, T ;

Loupatov, A ;

Munekata, E ;

Kaul, SC ;

Wadhwa, R ;

Imamura, T .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2001, 280 (04) :1203-1209

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