Deutsch
 
Benutzerhandbuch Datenschutzhinweis Impressum Kontakt
  DetailsucheBrowse

Datensatz

DATENSATZ AKTIONENEXPORT

Freigegeben

Zeitschriftenartikel

Expression of a soluble glycine binding domain of the NMDA receptor in Escherichia coli

MPG-Autoren
/persons/resource/persons118039

Betz,  H.
Neurochemistry Department, Max Planck Institute for Brain Research, Max Planck Society;

/persons/resource/persons208063

Kuhse,  J.
Neurochemistry Department, Max Planck Institute for Brain Research, Max Planck Society;

Externe Ressourcen
Es sind keine Externen Ressourcen verfügbar
Volltexte (frei zugänglich)
Es sind keine frei zugänglichen Volltexte verfügbar
Ergänzendes Material (frei zugänglich)
Es sind keine frei zugänglichen Ergänzenden Materialien verfügbar
Zitation

Neugebauer, R., Betz, H., & Kuhse, J. (2003). Expression of a soluble glycine binding domain of the NMDA receptor in Escherichia coli. Biochemical and Biophysical Research Communications, 305(3), 476-483.


Zitierlink: http://hdl.handle.net/11858/00-001M-0000-002E-1C0B-C
Zusammenfassung
Glycine is an essential co-agonist of the excitatory N-methyl-D-aspartate (NMDA) receptor. The glycine binding site of this subtype of ionotropic glutamate receptors is formed by the S1 and S2 regions of the NR1 subunit. Here, different S1S2 fusion proteins were expressed and purified from Escherichia coli cultures, and refolding protocols were established allowing the production of 30 mg of soluble S1S2 fusion protein from 1 liter bacterial culture. After affinity purification and renaturation, two of the fusion proteins (S1S2 and S1S2-V1) bound the competitive glycine site antagonist [H-3]MDL105,519 with K-d values of 9.35 and 3.9 nM, respectively. In contrast, with three other constructs (S1S2M, S1S2-V2, and -V3) saturable ligand binding could not be obtained. These results redefine the S1S2 domains required for high-affinity glycine binding. Furthermore, our high-affinity binding proteins may be used for the large-scale production of the glycine binding core region for future structural studies. (C) 2003 Elsevier Science (USA). All rights reserved.