Agonist binding to purified glycine receptor reconstituted into giant liposomes elicits two types of chloride currents

Abstract

Using 'inside-out' membrane patches obtained from reconstituted giant liposomes containing purified glycine receptor from rat spinal cord, we have detected chloride currents elicited in response to the presence of the agonists glycine or B-alanine. Regardless of the agonist employed, two different patterns of single channel currents could be detected, which differ in their main conductance, complexity of substates and opening frequency. In agreement with the expectations of glycine receptor heterogeneity suggested recently at the mRNA and cDNA level, our results indicate the existence of functionally different glycine receptors in the adult rat spinal cord.

Molecular and Electrophysiological Characterization of a Novel Cation Channel of Trypanosoma cruzi

“…Reconstitution of acetylcholine receptors [47], glycine receptors [48], glutamate receptors [49], KcsA [52], [59] and KirBac1.1 [57] indicate that the proteins are oriented “right-side out”, meaning with the intracellular side facing the bath. The direction of the rectification observed for TcCat (Fig.…”

Annexin 6 Modulates the Maxi-chloride Channel of the Apical Membrane of Syncytiotrophoblast Isolated from Human Placenta

“…Reconstitution of the Apical Membrane into Giant Liposomes-Giant liposomes were prepared by submitting a mixture of the isolated apical membrane vesicles and asolectin lipid vesicles to a partial dehydration/ rehydration cycle, as reported by Riquelme et al (25). An aliquot containing 100 -150 g of membrane protein was mixed with 2 ml of a 13 mM (in terms of lipid phosphorus) suspension of the asolectin vesicles.…”

Convulsants Acting at the Inhibitory Glycine Receptor

“…1) (GRAHAM et al 1981(GRAHAM et al , 1983(GRAHAM et al , 1985PFEIFFER et al 1982;BECKER et al 1986). Upon reconstitution in phospholipid vesicles, the isolated receptor mediates glycine-enhanced, strychnine-sensitive anion translocation (GARCIA-CALVO et al 1989;RIQUELME et al 1990). Upon reconstitution in phospholipid vesicles, the isolated receptor mediates glycine-enhanced, strychnine-sensitive anion translocation (GARCIA-CALVO et al 1989;RIQUELME et al 1990).…”

“…A sequence comparison shows that this region of the acetylcholine receptor corresponds to the pre-M1 domain of the highly homologous glycine receptor (BETZ 1990). In fact, analysis of proteolytic fragments of the glycine receptor has shown that UV illumination leads to incorporation of [3H]strychnine into peptide fragments of this ai-subunit region (RUIZ-GOMEZ et al 1990). In fact, analysis of proteolytic fragments of the glycine receptor has shown that UV illumination leads to incorporation of [3H]strychnine into peptide fragments of this ai-subunit region (RUIZ-GOMEZ et al 1990).…”