Protocol

Structural Proteomics

Volume 1261 of the series Methods in Molecular Biology pp 91-114

Date:

The Production of Multiprotein Complexes in Insect Cells Using the Baculovirus Expression System

  • Wassim AbdulrahmanAffiliated withInstitut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/UDSFriedrich Miescher Institute for Biomedical Research
  • , Laura RaduAffiliated withInstitut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/UDS
  • , Frederic GarzoniAffiliated withEuropean Molecular Biology Laboratory (EMBL), Grenoble Outstation and Unit of Virus Host-Cell Interactions (UVHCI), Universite Grenoble Alpes-EMBL-CNRS, UMR 5233
  • , Olga KolesnikovaAffiliated withInstitut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/UDS
  • , Kapil GuptaAffiliated withEuropean Molecular Biology Laboratory (EMBL), Grenoble Outstation and Unit of Virus Host-Cell Interactions (UVHCI), Universite Grenoble Alpes-EMBL-CNRS, UMR 5233
  • , Judit Osz-PapaiAffiliated withInstitut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/UDS
  • , Imre BergerAffiliated withEuropean Molecular Biology Laboratory (EMBL), Grenoble Outstation and Unit of Virus Host-Cell Interactions (UVHCI), Universite Grenoble Alpes-EMBL-CNRS, UMR 5233
  • , Arnaud PoterszmanAffiliated withInstitut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/UDS Email author 

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Abstract

The production of a homogeneous protein sample in sufficient quantities is an essential prerequisite not only for structural investigations but represents also a rate-limiting step for many functional studies. In the cell, a large fraction of eukaryotic proteins exists as large multicomponent assemblies with many subunits, which act in concert to catalyze specific activities. Many of these complexes cannot be obtained from endogenous source material, so recombinant expression and reconstitution are then required to overcome this bottleneck. This chapter describes current strategies and protocols for the efficient production of multiprotein complexes in large quantities and of high quality, using the baculovirus/insect cell expression system.

Key words

Recombinant protein complex production BEVS Baculovirus Insect cells Infection and coinfection methods Multigene expression Multiprotein complex