Isolation and partial characterization of calcium-activated chloride ion channels from thylakoids.
The goal of the present work was to characterize the thylakoid anion channel. Toward this purpose a new, more sensitive, assay was developed to measure chloride channel activity of solubilized and partially purified thylakoid membrane proteins. In this assay potassium channel function was reduced or eliminated by the use of cesium as the coion of chloride. Under these conditions calcium but not magnesium stimulated anion uptake, suggesting the presence of a calcium-activated anion channel in isolated thylakoid membrane proteins. A polyclonal antibody raised against an anion channel isolated from epithelial membranes immunoreacted with three thylakoid membrane proteins; a doublet in the 62 to 66-kDa range and a single band in the 57 to 59-kDa region suggesting that the thylakoid anion channel may be homologous to the mammalian chloride channel. Furthermore, the cDNA which encodes the epithelial chloride channel hybridized on Northern blots prepared from corn total RNA with three mRNA bands.
Department of Biochemistry, West Virginia University School of Medicine, Morgantown 26506.
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D001692:Biological Transport; D002118:Calcium / Q000494:pharmacology*; D018118:Chloride Channels / Q000187:drug effects / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism*; D002736:Chloroplasts / Q000737:chemistry* / Q000276:immunology; D007425:Intracellular Membranes / Q000737:chemistry* / Q000276:immunology; D009693:Nucleic Acid Hybridization; D012689:Sequence Homology, Nucleic Acid; D014675:Vegetables
D018118:Chloride Channels; D002712:Chlorides; D002118:Calcium