title: Co-expression of protein complexes in prokaryotic and eukaryotic hosts: experimental procedures, database tracking and case studies.
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.
authors.affiliation: Institut de Génétique et Biologie Moléculaire et Cellulaire, 1 Rue Laurent Fries, BP 163, Illkirch CEDEX, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Romier
authors.forename: Marouane
authors.initials: M
authors.lastname: Ben Jelloul
authors.forename: Shira
authors.initials: S
authors.lastname: Albeck
authors.forename: Gretel
authors.initials: G
authors.lastname: Buchwald
authors.forename: Didier
authors.initials: D
authors.lastname: Busso
authors.forename: Patrick H N
authors.initials: PH
authors.lastname: Celie
authors.forename: Evangelos
authors.initials: E
authors.lastname: Christodoulou
authors.forename: Valeria
authors.initials: V
authors.lastname: De Marco
authors.forename: Suzan
authors.initials: S
authors.lastname: van Gerwen
authors.forename: Puck
authors.initials: P
authors.lastname: Knipscheer
authors.forename: Joyce H
authors.initials: JH
authors.lastname: Lebbink
authors.forename: Valerie
authors.initials: V
authors.lastname: Notenboom
authors.forename: Arnaud
authors.initials: A
authors.lastname: Poterszman
authors.forename: Natacha
authors.initials: N
authors.lastname: Rochel
authors.forename: Serge X
authors.initials: SX
authors.lastname: Cohen
authors.forename: Tamar
authors.initials: T
authors.lastname: Unger
authors.forename: Joel L
authors.initials: JL
authors.lastname: Sussman
authors.forename: Dino
authors.initials: D
authors.lastname: Moras
authors.forename: Titia K
authors.initials: TK
authors.lastname: Sixma
authors.forename: Anastassis
authors.initials: A
authors.lastname: Perrakis
mesh_terms: D000465:Algorithms; D000818:Animals; D016494:Computer Security; D003198:Computer Simulation; D018844:Cyclin-Dependent Kinases / Q000378:metabolism; D004260:DNA Repair; D030541:Databases, Genetic; D004926:Escherichia coli / Q000378:metabolism; D005057:Eukaryotic Cells / Q000378:metabolism*; D005822:Genetic Vectors; D019451:Information Management; D007313:Insecta / Q000378:metabolism; D011387:Prokaryotic Cells / Q000378:metabolism*; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis*; D035362:Transcription Factor TFIID / Q000235:genetics; D044767:Ubiquitin-Protein Ligases / Q000235:genetics; D000095043:Cyclin-Dependent Kinase-Activating Kinase
chemical_list: D018160:Receptors, Cytoplasmic and Nuclear; D011994:Recombinant Proteins; D035362:Transcription Factor TFIID; D012313:RNA; D044767:Ubiquitin-Protein Ligases; D018844:Cyclin-Dependent Kinases; D000095043:Cyclin-Dependent Kinase-Activating Kinase