Eukaryotic expression: developments for structural proteomics.
Journal article, 2006

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), baculovirus-infected 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 co-expression, selenomethionine labelling (in all three eukaryotic systems) and control of glycosylation (for secreted proteins in mammalian cells) are assessed.

Molecular

Cloning

Animals

Selenomethionine

Proteomics

metabolism

genetics

Gene Expression

Cultured

Eukaryotic Cells

methods

Glycosylation

Yeasts

Cells

Baculoviridae

diagnostic use

metabolism

Author

A R Aricescu

Wellcome Trust Centre for Human Genetics

R Assenberg

Wellcome Trust Centre for Human Genetics

Joakim Norbeck

Chalmers

Aston University

D Busso

IGBMC Institut de Genetique et de Biologie Moleculaire et Cellulaire

V T Chang

John Radcliffe Hospital

S J Davis

John Radcliffe Hospital

A Dubrovsky

Karolinska Institutet

Lena Gustafsson

Chalmers, Chemical and Biological Engineering

Kristina Hedfalk

Chalmers, Chemical and Biological Engineering, Molecular Biotechnology

U Heinemann

Max Delbruck Center for Molecular Medicine

I M Jones

University of Reading

D Ksiazek

Max Planck Institute

C Lang

Technische Universität Berlin

K Maskos

Max Planck Institute

A Messerschmidt

Max Planck Institute

S Macieira

Max Planck Institute

Y Peleg

Weizmann Institute of Science

A Perrakis

The Netherlands Cancer Institute

A Poterszman

IGBMC Institut de Genetique et de Biologie Moleculaire et Cellulaire

G Schneider

John Radcliffe Hospital

T K Sixma

The Netherlands Cancer Institute

J L Sussman

Weizmann Institute of Science

G Sutton

Wellcome Trust Centre for Human Genetics

N Tarboureich

Institut Laue-Langevin

T Zeev-Ben-Mordehai

Weizmann Institute of Science

E Yvonne Jones

Wellcome Trust Centre for Human Genetics

Acta Crystallographica Section D: Biological Crystallography

0907-4449 (ISSN) 1399-0047 (eISSN)

Vol. 62 Pt 10 1114-24

Subject Categories

Biological Sciences

DOI

10.1107/S0907444906029805

PubMed

17001089

More information

Latest update

9/10/2018