A novel and rapid approach to isolating functional ryanodine receptors.
Conventional methods of isolating and reconstituting ryanodine receptors (RyRs) from native membranes into proteoliposomes take a minimum of 2 days to complete. We have developed an alternative strategy that can be used to isolate and reconstitute functional RyRs in just 3 h with a similar degree of purification. RyRs isolated by this method display characteristic functional behaviour as assessed by radioligand binding and single channel analyses.
West
Duncan J
DJ
Department of Cardiac Medicine, National Heart and Lung Institute, Faculty of Medicine, Imperial College of Science, Technology and Medicine, Dovehouse Street, London SW3 6LY, UK.
Smith
Eileen C J
EC
Williams
Alan J
AJ
D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects / Q000502:physiology; D015153:Blotting, Western; D002118:Calcium / Q000494:pharmacology; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D046911:Macromolecular Substances; D008564:Membrane Potentials / Q000502:physiology; D009206:Myocardium / Q000737:chemistry; D011510:Proteolipids / Q000737:chemistry / Q000302:isolation & purification; D011869:Radioligand Assay; D012433:Ryanodine / Q000737:chemistry; D019837:Ryanodine Receptor Calcium Release Channel / Q000737:chemistry / Q000302:isolation & purification*; D012519:Sarcoplasmic Reticulum / Q000737:chemistry; D012756:Sheep; D012995:Solubility; D014316:Tritium
D046911:Macromolecular Substances; D011510:Proteolipids; D019837:Ryanodine Receptor Calcium Release Channel; C020485:proteoliposomes; D014316:Tritium; D012433:Ryanodine; D002118:Calcium