Expression in animal cells of the 5-HT1A receptor by a vaccinia virus vector system

FEBS Lett. 1992 Apr 27;301(3):303-6. doi: 10.1016/0014-5793(92)80262-f.

Abstract

The co-infection or infection-transfection variants of the T7 RNA polymerase/vaccinia vector system were used to express 5-HT1ARs in COS-7, BSC-40 and GH3 cells, with co-infection giving ca. 3-fold higher level than infection-transfection. Binding affinities were similar to those of the endogenous 5-HT1AR, with highest affinities for 5-HT and 8-OH-DPAT. Functional properties were demonstrated by assays of agonist-stimulated GTPase activity and its inhibition by pertussin toxin. Immunoblot assays showed expression of the unglycosylated and glycosylated receptor protein in the membrane and, surprisingly, in the cytosolic fractions.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 8-Hydroxy-2-(di-n-propylamino)tetralin
  • Animals
  • Binding, Competitive
  • Blotting, Western
  • Cell Line
  • Cell Membrane / metabolism
  • DNA-Directed RNA Polymerases / metabolism
  • GTP Phosphohydrolases / metabolism
  • Gene Expression
  • Genes, Viral
  • Genetic Vectors*
  • Receptors, Serotonin / genetics*
  • Receptors, Serotonin / metabolism
  • Serotonin / metabolism
  • T-Phages / enzymology
  • Tetrahydronaphthalenes / metabolism
  • Vaccinia virus / genetics*

Substances

  • Receptors, Serotonin
  • Tetrahydronaphthalenes
  • Serotonin
  • 8-Hydroxy-2-(di-n-propylamino)tetralin
  • DNA-Directed RNA Polymerases
  • GTP Phosphohydrolases