Co-expression of protein complexes in prokaryotic and eukaryotic hosts: experimental procedures, database tracking
Christophe Romier1, Marouane Ben Jelloul, Shira Albeck
1Institut de Génétique et Biologie Moléculaire et Cellulaire, 1 Rue Laurent Fries, BP 163, Illkirch CEDEX, France.
Abstract:
Structure determination and functional characterization of macromolecular complexes requires the purification of the different subunits in large quantities and their assembly into a functional entity. Although isolation and structure determination of endogenous complexes has been reported, much progress has to be made to make this technology easily accessible. Co-expression of subunits within hosts such as Escherichia coli and insect cells has become more and more amenable, even at the level of high-throughput projects. As part of SPINE (Structural Proteomics In Europe), several laboratories have investigated the use co-expression techniques for their projects, trying to extend from the common binary expression to the more complicated multi-expression systems. A new system for multi-expression in E. coli and a database system dedicated to handle co-expression data are described. Results are also reported from various case studies investigating different methods for performing co-expression in E. coli and insect cells.
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