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

Christophe Romier , Marouane Ben Jelloul , Shira Albeck , Gretel Buchwald , Didier Busso (1) , Patrick H N Celie , Evangelos Christodoulou , Valeria de Marco , Suzan van Gerwen , Puck Knipscheer , Joyce H Lebbink , Valerie Notenboom , Arnaud Poterszman (1) , Natacha Rochel (1) , Serge X Cohen , Tamar Unger , Joel L Sussman , Dino Moras (1) , Titia K Sixma , Anastassis Perrakis

1 IGBMC - Institut de génétique et biologie moléculaire et cellulaire

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.

Domaines: Biologie moléculaire

Christophe Romier, Marouane Ben Jelloul, Shira Albeck, Gretel Buchwald, Didier Busso, et al.. Co-expression of protein complexes in prokaryotic and eukaryotic hosts: experimental procedures, database tracking and case studies.. Acta crystallographica Section D : Structural biology [1993-..], 2006, 62 (Pt 10), pp.1232-42. ⟨10.1107/S0907444906031003⟩. ⟨hal-00188127⟩