Structural Proteomics pp 91-114

Part of the Methods in Molecular Biology book series (MIMB, volume 1261)

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

  • Wassim Abdulrahman
  • Laura Radu
  • Frederic Garzoni
  • Olga Kolesnikova
  • Kapil Gupta
  • Judit Osz-Papai
  • Imre Berger
  • Arnaud Poterszman
Protocol

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 

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Wassim Abdulrahman
    • 1
    • 2
  • Laura Radu
    • 1
  • Frederic Garzoni
    • 3
  • Olga Kolesnikova
    • 1
  • Kapil Gupta
    • 3
  • Judit Osz-Papai
    • 1
  • Imre Berger
    • 3
  • Arnaud Poterszman
    • 1
  1. 1.Institut de Génétique et de Biologie Moléculaire et CellulaireCNRS/INSERM/UDSIllkirch CedexFrance
  2. 2.Friedrich Miescher Institute for Biomedical ResearchBaselSwitzerland
  3. 3.European Molecular Biology Laboratory (EMBL), Grenoble Outstation and Unit of Virus Host-Cell Interactions (UVHCI)Universite Grenoble Alpes-EMBL-CNRS, UMR 5233Grenoble Cedex 9France

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