Co-expression of protein complexes in prokaryotic and eukaryotic hosts: experimental procedures, database tracking and case studies.

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Romier, Christophe | Ben Jelloul, Marouane | Albeck, Shira | Buchwald, Gretel | Busso, Didier | Celie, Patrick H N | Christodoulou, Evangelos | de Marco, Valeria | van Gerwen, Suzan | Knipscheer, Puck | Lebbink, Joyce H | Notenboom, Valerie | Poterszman, Arnaud | Rochel, Natacha | Cohen, Serge X | Unger, Tamar | Sussman, Joel L | Moras, Dino | Sixma, Titia K | Perrakis, Anastassis

Edité par CCSD ; International Union of Crystallography -

International audience. 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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