Recovery of homogeneous and functional beta 2-adrenergic receptors from extracellular baculovirus particles

Expression in baculovirus-infected insect cells allows sufficient production of G-protein coupled receptor for structural studies. An important drawback of this expression system comes from the presence of unprocessed and biologically inactive receptors that have to be eliminated during receptor pur...

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Published inNature biotechnology Vol. 15; no. 12; pp. 1300 - 1304
Main Authors Loisel, T P, Ansanay, H, St-Onge, S, Gay, B, Boulanger, P, Strosberg, A D, Marullo, S, Bouvier, M
Format Journal Article
LanguageEnglish
Published United States 01.11.1997
Subjects
Adenylyl Cyclases - metabolism
Animals
Baculoviridae - genetics
Cell Line
Humans
Microscopy, Immunoelectron
Protein Binding
Protein Processing, Post-Translational
Receptors, Adrenergic, beta-2 - genetics
Receptors, Adrenergic, beta-2 - metabolism
Receptors, Adrenergic, beta-2 - physiology
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
Spodoptera
Virion - genetics
Virion - ultrastructure
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Summary:Expression in baculovirus-infected insect cells allows sufficient production of G-protein coupled receptor for structural studies. An important drawback of this expression system comes from the presence of unprocessed and biologically inactive receptors that have to be eliminated during receptor purification steps. We show that viral particles released from Sf9 cells infected with a recombinant baculovirus coding for the human beta 2-adrenergic receptor (beta 2AR) cDNA contain glycosylated and biologically active beta 2AR. In addition, post-translational modifications known to modulate receptor activity were found to occur in these particles.
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ISSN:1087-0156