Pharmacological characterization and immunoaffinity purification of metabotropic glutamate receptor from Drosophila overexpressed in Sf9 cells.

Published

Journal Article

Metabotropic glutamate receptors (mGluRs) play important roles in the function and regulation of the central nervous system. Structural studies are necessary for the detailed understanding of their mechanisms of action. However, overexpression and purification of functional receptors in quantities required for these studies proves to be a major challenge. In this study we report the overexpression of a Drosophila melanogaster mGluR (DmGluRA) by using a baculovirus-insect cell expression system. Expression was tested in two different insect cell hosts (Sf9 and Hi5) and analyzed by performing expression kinetics. Pharmacological characterization of the recombinant receptor by radioactive glutamate binding assays showed a profile similar to group II mGluRs, as previously reported, when the receptor was expressed in mammalian systems. The B(max) value reached 11 pM receptor/mg Sf9-membrane protein. A monoclonal antibody against DmGluRA was generated by genetic immunization and used to purify the receptor.

Full Text

Duke Authors

Cited Authors

  • Panneels, V; Eroglu, C; Cronet, P; Sinning, I

Published Date

  • August 2003

Published In

Volume / Issue

  • 30 / 2

Start / End Page

  • 275 - 282

PubMed ID

  • 12880777

Pubmed Central ID

  • 12880777

International Standard Serial Number (ISSN)

  • 1046-5928

Language

  • eng

Conference Location

  • United States