Proteomic analysis of complexes formed by human topoisomerase I

被引：42

作者：

Czubaty, A ^{[1
]}

Girstun, A ^{[1
]}

Kowalska-Loth, B ^{[1
]}

Trzcinska, AA ^{[1
]}

Purta, E ^{[1
]}

Winczura, A ^{[1
]}

Grajkowski, W ^{[1
]}

Staron, K ^{[1
]}

机构：

[1] Warsaw Univ, Inst Biochem, PL-02096 Warsaw, Poland

来源：

BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS | 2005年 / 1749卷 / 01期

关键词：

proteomics; topoisomerase I; protein partners; Co-immunoprecipitation; affinity chromatography;

D O I：

10.1016/j.bbapap.2005.03.007

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：

Human topoisomerase I is a nuclear enzyme that catalyses DNA relaxation and phosphorylation of SR proteins. Topoisomerase I participates in several protein-protein interactions. We performed a proteomic analysis of protein partners of topoisomerase 1. Two methods were applied to proteins of the nuclear extract of HeLa cells: a co-immunoprecipitation and an affinity chromatography combined with mass spectrometry. Complexes formed by topoisomerase I with its protein partners were immunoprecipitated by scleroderma anti-topoisomerase I antibodies. To identify binding sites for the protein partners, baits corresponding to fragments of topoisomerase I were constructed and used in the affinity chromatography. The N-terminal domain and the cap region of the core domain appeared to be the main regions that bound proteins. We identified 36 nuclear proteins that were associated with topoisomerase 1. The proteins were mainly involved in RNA metabolism. We found 29 new and confirmed 7 previously identified protein partners of topoisomerase 1. More than 40% proteins that associate with the cap region contain two closely spaced RRM domains. Docking calculations identified the RRM domains as a possible site for the interaction of these proteins with the cap region. (c) 2005 Elsevier B.V All rights reserved.

引用

页码：133 / 141

页数：9

共 57 条

[1] The human U5-220kD protein (hPrp8) forms a stable RNA-free complex with several US-specific proteins, including an RNA unwindase, a homologue of ribosomal elongation factor EF-2, and a novel WD-40 protein [J].