Purification of G protein subunits from Sf9 insect cells using hexahistidine-tagged alpha and beta gamma subunits

Methods Mol Biol. 2004;237:21-38. doi: 10.1385/1-59259-430-1:21.

Abstract

G protein-mediated pathways are the most fundamental mechanisms of cell signaling. In order to analyze these pathways, the availability of purified recombinant G proteins are critically important. Using Sf9-Baculovirus expression system, a general and simplified method to purify various G protein subunits is described in this chapter. This method is useful for purification of most of G protein subunits.

Publication types

  • Review

MeSH terms

  • Animals
  • Baculoviridae / genetics
  • Cell Line
  • Chromatography / methods*
  • GTP-Binding Protein alpha Subunits / genetics
  • GTP-Binding Protein alpha Subunits / isolation & purification
  • GTP-Binding Protein beta Subunits / genetics
  • GTP-Binding Protein beta Subunits / isolation & purification
  • GTP-Binding Protein gamma Subunits / genetics
  • GTP-Binding Protein gamma Subunits / isolation & purification
  • Heterotrimeric GTP-Binding Proteins / genetics
  • Heterotrimeric GTP-Binding Proteins / isolation & purification*
  • Histidine / metabolism
  • Insecta
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification*
  • Signal Transduction

Substances

  • GTP-Binding Protein alpha Subunits
  • GTP-Binding Protein beta Subunits
  • GTP-Binding Protein gamma Subunits
  • Recombinant Proteins
  • Histidine
  • Heterotrimeric GTP-Binding Proteins