Co-expression of protein complexes in prokaryotic and eukaryotic hosts: experimental procedures, database tracking and case studies.
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.
Romier
Christophe
C
Institut de Génétique et Biologie Moléculaire et Cellulaire, 1 Rue Laurent Fries, BP 163, Illkirch CEDEX, France.
Ben Jelloul
Marouane
M
Albeck
Shira
S
Buchwald
Gretel
G
Busso
Didier
D
Celie
Patrick H N
PH
Christodoulou
Evangelos
E
De Marco
Valeria
V
van Gerwen
Suzan
S
Knipscheer
Puck
P
Lebbink
Joyce H
JH
Notenboom
Valerie
V
Poterszman
Arnaud
A
Rochel
Natacha
N
Cohen
Serge X
SX
Unger
Tamar
T
Sussman
Joel L
JL
Moras
Dino
D
Sixma
Titia K
TK
Perrakis
Anastassis
A
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
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