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PMID: 10423421 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Investigation of the alpha(1)-glycine receptor channel-opening kinetics in the submillisecond time domain.

Biophysical journal ·Vol. 77 ·No. 2 ·1999-08-00 ·Pages 727-38

Grewer C

Abstract

The activation and desensitization kinetics of the human alpha(1)-homooligomeric glycine receptor, which was transiently expressed in HEK 293 cells, were studied with a 100-microseconds time resolution to determine the rate and equilibrium constants of individual receptor reaction steps. Concentration jumps of the activating ligands glycine and beta-alanine were initiated by photolysis of caged, inactive precursors and were followed by neurotransmitter binding, receptor-channel opening, and receptor desensitization steps that were separated along the time axis. Analysis of the ligand concentration-dependence of these processes allows the determination of 1) the rate constants of glycine binding, k(+1) approximately 10(7) M(-1) s(-1), and dissociation, k(-1) = 1900 s(-1); 2) the rates of receptor-channel opening, k(op) = 2200 s(-1), and closing, k(cl) = 38 s(-1); 3) the receptor desensitization rate, alpha = 0.45 s(-1); 4) the number of occupied ligand binding sites necessary for receptor-channel activation and desensitization, n >/= 3; and 5) the maximum receptor-channel open probability, p(0) > 0.95. The kinetics of receptor-channel activation are insensitive to the transmembrane potential. A general model for glycine receptor activation explaining the experimental data consists of a sequential mechanism based on rapid ligand-binding steps preceding a rate-limiting receptor-channel opening reaction and slow receptor desensitization.

MeSH Terms
Binding Sites Biophysical Phenomena Biophysics Cell Line Electrochemistry Glycine/metabolism Humans Kinetics Ligands Membrane Potentials Receptors, Glycine/genetics,metabolism Recombinant Proteins/genetics,metabolism beta-Alanine/metabolism
Chemicals
Ligands Receptors, Glycine Recombinant Proteins beta-Alanine Glycine
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Grewer C
Max-Planck-Institut for Biophysics, D-60596 Frankfurt, Germany. grewer@kennedy.biophys.mpg.de
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Article Info
Journal
Biophysical journal
Abbr.
Biophys J
ISSN
0006-3495
DOI
10.1016/S0006-3495(99)76927-4
Email
grewer@kennedy.biophys.mpg.de
Published
1999-08-00
Pages
727-38
Language
English
Region
United States
NLM ID
0370626
PMCID
PMC1300367
Subset
IM
External Links
PubMed source DOI: 10.1016/S0006-3495(99)76927-4 PMC Full Text View journal details
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