title: The multiBac protein complex production platform at the EMBL.
abstract: Proteomics research revealed the impressive complexity of eukaryotic proteomes in unprecedented detail. It is now a commonly accepted notion that proteins in cells mostly exist not as isolated entities but exert their biological activity in association with many other proteins, in humans ten or more, forming assembly lines in the cell for most if not all vital functions.(1,2) Knowledge of the function and architecture of these multiprotein assemblies requires their provision in superior quality and sufficient quantity for detailed analysis. The paucity of many protein complexes in cells, in particular in eukaryotes, prohibits their extraction from native sources, and necessitates recombinant production. The baculovirus expression vector system (BEVS) has proven to be particularly useful for producing eukaryotic proteins, the activity of which often relies on post-translational processing that other commonly used expression systems often cannot support.(3) BEVS use a recombinant baculovirus into which the gene of interest was inserted to infect insect cell cultures which in turn produce the protein of choice. MultiBac is a BEVS that has been particularly tailored for the production of eukaryotic protein complexes that contain many subunits.(4) A vital prerequisite for efficient production of proteins and their complexes are robust protocols for all steps involved in an expression experiment that ideally can be implemented as standard operating procedures (SOPs) and followed also by non-specialist users with comparative ease. The MultiBac platform at the European Molecular Biology Laboratory (EMBL) uses SOPs for all steps involved in a multiprotein complex expression experiment, starting from insertion of the genes into an engineered baculoviral genome optimized for heterologous protein production properties to small-scale analysis of the protein specimens produced.(5-8) The platform is installed in an open-access mode at EMBL Grenoble and has supported many scientists from academia and industry to accelerate protein complex research projects.
authors.affiliation: iberger@embl.fr
authors.forename: Imre
authors.initials: I
authors.lastname: Berger
authors.forename: Frederic
authors.initials: F
authors.lastname: Garzoni
authors.forename: Maxime
authors.initials: M
authors.lastname: Chaillet
authors.forename: Matthias
authors.initials: M
authors.lastname: Haffke
authors.forename: Kapil
authors.initials: K
authors.lastname: Gupta
authors.forename: Alice
authors.initials: A
authors.lastname: Aubert
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D008967:Molecular Biology / Q000295:instrumentation / Q000379:methods / Q000592:standards; D046912:Multiprotein Complexes / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D061987:Sf9 Cells / Q000378:metabolism / Q000821:virology*; D018411:Spodoptera
chemical_list: D046912:Multiprotein Complexes; D011994:Recombinant Proteins
references.citation: Nat Biotechnol. 2004 Dec;22(12):1583-7
references.pmid: 15568020
references.citation: Nat Methods. 2009 Jun;6(6):447-50
references.pmid: 19412171
references.citation: Nat Biotechnol. 2005 May;23(5):567-75
references.pmid: 15877075
references.citation: Nat Methods. 2006 Dec;3(12):1021-32
references.pmid: 17117155
references.citation: Protein Expr Purif. 2009 Jun;65(2):122-32
references.pmid: 19174191
references.citation: Nature. 2011 Apr 28;472(7344):448-53
references.pmid: 21525927
references.citation: J Struct Biol. 2011 Aug;175(2):198-208
references.pmid: 21419851
references.citation: Science. 2000 Apr 28;288(5466):640-9
references.pmid: 10784442
references.citation: Crit Rev Biochem Mol Biol. 2006 May-Jun;41(3):105-78
references.pmid: 16858867
references.citation: Trends Biochem Sci. 2012 May;37(5):189-98
references.pmid: 22436288
references.citation: Nat Struct Mol Biol. 2012 Jun 05;19(6):560-7
references.pmid: 22664983
references.citation: Nature. 2012 Mar 21;484(7393):208-13
references.pmid: 22437499
references.citation: Curr Protoc Protein Sci. 2008 Feb;Chapter 5:Unit 5.20
references.pmid: 18429060
references.citation: Nature. 2007 Dec 13;450(7172):973-82
references.pmid: 18075576
references.citation: Trends Biochem Sci. 2012 Feb;37(2):49-57
references.pmid: 22154230
references.citation: J Struct Biol. 2010 Oct;172(1):45-54
references.pmid: 20178849
references.citation: Curr Genomics. 2009 Dec;10(8):558-72
references.pmid: 20514218