English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  High-level expression of functional glutamate receptor channels in insect cells

Keinänen, K., Köhr, G., Seeburg, P. H., Laukkanen, M., & Oker−Blom, C. (1994). High-level expression of functional glutamate receptor channels in insect cells. Nature Biotechnology, 12(8), 804-806. doi:10.1038/nbt0894-802.

Item is

Files

show Files
hide Files
:
NatBiotechnol_12_1994_802.pdf (Any fulltext), 595KB
 
File Permalink:
-
Name:
NatBiotechnol_12_1994_802.pdf
Description:
-
Visibility:
Restricted (Max Planck Institute for Medical Research, MHMF; )
MIME-Type / Checksum:
application/pdf
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-

Locators

show
hide
Description:
-
Locator:
https://doi.org/10.1038/nbt0894-802 (Any fulltext)
Description:
-

Creators

show
hide
 Creators:
Keinänen, Kari, Author
Köhr, Georg1, 2, 3, Author              
Seeburg, Peter H.1, Author              
Laukkanen, Marja−Leena, Author
Oker−Blom, Christian, Author
Affiliations:
1Department of Molecular Neurobiology, Max Planck Institute for Medical Research, Max Planck Society, ou_1497704              
2Directly responsible to the Managing Director, Max Planck Institute for Medical Research, Max Planck Society, ou_persistent22              
3Georg Köhr Group, Max Planck Institute for Medical Research, Max Planck Society, ou_1497714              

Content

show
hide
Free keywords: -
 Abstract: We have expressed glutamate-gated ion channels in Spodoptera frugiperda Sf21 insect cells using a recombinant baculovirus system. Cells infected with recombinant baculoviruses encoding the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA)-selective glutamate receptor channel subunits GluR-B and GluR-D displayed specific high-affinity [3H]AMPA binding (apparent dissociation constant Kd of 15 nM for GluR-B and 40 nM for GluR-D) with pharmacological profiles typical of AMPA receptors. The binding reached maximal levels (Bmax of 15-30 pmol per mg of membrane protein) by 3-4 days postinfection. AMPA, glutamate and kainate triggered inward currents in GluR expressing cells, indicating assembly of functional homomeric channels. Formation of heteromeric GluR-B/D channels in doubly-infected cells was evident from the diagnostic current-voltage relations of AMPA-activated whole-cell currents. For the solubilization of the receptor, nonionic detergents Triton X-100, n-octyl-D-glucoside and n-dodecylmaltoside proved most effective. Detergent-solubilized receptor preparations were stable, retained their characteristic ligand-binding properties and bound to immobilized wheat germ lectin, demonstrating the glycosylation of insect cell-expressed GluR subunits. The expression level of 300-400 micrograms of receptor protein per liter of suspension culture should facilitate production of glutamate receptors for biochemical and structural studies.

Details

show
hide
Language(s): eng - English
 Dates: 1994-02-161994-05-091994
 Publication Status: Published in print
 Pages: 5
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 666772
DOI: 10.1038/nbt0894-802
URI: https://www.ncbi.nlm.nih.gov/pubmed/7519022
Other: 4079
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Nature Biotechnology
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: New York : Gale Group Inc.
Pages: - Volume / Issue: 12 (8) Sequence Number: - Start / End Page: 804 - 806 Identifier: ISSN: 1087-0156
CoNE: https://pure.mpg.de/cone/journals/resource/954926980065