Eukaryotic expression: developments for structural proteomics

被引：73

作者：

Aricescu, A. R.

Assenberg, R.

Bill, R. M.

Busso, D.

Chang, V. T.

Davis, S. J.

Dubrovsky, A.

Gustafsson, L.

Hedfalk, K.

Heinemann, U.

Jones, I. M.

Ksiazek, D.

Lang, C.

Maskos, K.

Messerschmidt, A.

Macieira, S.

Peleg, Y.

Perrakis, A.

Poterszman, A.

Schneider, G.

Sixma, T. K.

Sussman, J. L.

Sutton, G.

Tarboureich, N.

Zeev-Ben-Mordehai, T.

Jones, E. Yvonne

机构：

[1] Wellcome Trust Ctr Human Genet, Div Struct Biol, Oxford OX3 7BN, England

[2] Wellcome Trust Ctr Human Genet, Oxford Prot Prod Facil, Oxford OX3 7BN, England

[3] Chalmers Univ Technol, Dept Chem & Biosci, SE-40530 Gothenburg, Sweden

[4] Inst Genet & Biol Mol & Cellulaire, F-67404 Illkirch Graffenstaden, France

[5] John Radcliffe Hosp, Weatherall Inst Mol Med, Oxford OX3 9DU, England

[6] Karolinska Inst, Dept Med Biochem & Biophys, SE-10951 Stockholm, Sweden

[7] Max Delbruck Ctr Mol Med, Dept Crystallog, D-13125 Berlin, Germany

[8] Univ Reading, Sch Biol Sci, Reading RG6 6AH, Berks, England

[9] Max Planck Inst Biochem, Dept Proteom & Signal Transduct, D-82152 Martinsried, Germany

[10] Tech Univ Berlin, Inst Biotechnol, D-13355 Berlin, Germany

[11] Weizmann Inst Sci, Israel Struct Proteom Ctr, Dept Biol Struct, IL-76100 Rehovot, Israel

[12] Netherlands Canc Inst, Div Mol Carcinogenisis, NL-1066 CX Amsterdam, Netherlands

[13] ILL Grenoble, EMBL Grenoble, F-38042 Grenoble 9, France

来源：

ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY | 2006年 / 62卷

基金：

英国医学研究理事会;

关键词：

D O I：

10.1107/S0907444906029805

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The production of sufficient quantities of protein is an essential prelude to a structure determination, but for many viral and human proteins this cannot be achieved using prokaryotic expression systems. Groups in the Structural Proteomics In Europe ( SPINE) consortium have developed and implemented high- throughput ( HTP) methodologies for cloning, expression screening and protein production in eukaryotic systems. Studies focused on three systems: yeast ( Pichia pastoris and Saccharomyces cerevisiae), baculovirusinfected insect cells and transient expression in mammalian cells. Suitable vectors for HTP cloning are described and results from their use in expression screening and protein-production pipelines are reported. Strategies for coexpression, selenomethionine labelling ( in all three eukaryotic systems) and control of glycosylation ( for secreted proteins in mammalian cells) are assessed.

引用

页码：1114 / 1124

页数：11

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