Sensitive procedures for measuring chloride fluxes mediated by the purified glycine receptor incorporated into phospholipid vesicles.
Two novel methods have been developed to directly measure chloride influx into purified glycine receptor-containing phospholipid vesicles. Using a method based on the fluorescence quenching of a chloride-sensitive probe entrapped into the vesicles, a chloride influx was detected which could be enhanced by glycine and completely abolished by the antagonist strychnine. In addition, by tracing the 36Cl- influx into the proteoliposomes, a stimulatory effect of both glycine and beta-alanine could be seen, which can be inhibited by strychnine and other glycine antagonists. These data, together with a previous report demonstrating ligand-mediated iodide fluxes in the same preparation (Biochemistry, 28 (1989) 6405-6409), clearly demonstrate the utility of the reconstituted receptor preparation to investigate some ion channel and pharmacological properties of the glycine receptor.
García-Calvo
M
M
Departamento de Biología Molecular UAM-CSIC, Facultad de Ciencias, Universidad Autónoma de Madrid, Spain.
Valdivieso
F
F
Mayor
F
F
Vázquez
J
J
D000818:Animals; D018118:Chloride Channels; D002712:Chlorides / Q000378:metabolism*; D005998:Glycine / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D015640:Ion Channel Gating / Q000187:drug effects; D008081:Liposomes; D008565:Membrane Proteins / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D018009:Receptors, Glycine; D017981:Receptors, Neurotransmitter / Q000187:drug effects / Q000378:metabolism*; D013331:Strychnine / Q000494:pharmacology; D015091:beta-Alanine / Q000494:pharmacology
D018118:Chloride Channels; D002712:Chlorides; D008081:Liposomes; D008565:Membrane Proteins; D018009:Receptors, Glycine; D017981:Receptors, Neurotransmitter; D015091:beta-Alanine; D013331:Strychnine; D005998:Glycine