title: Regulation of ryanodine receptors by calsequestrin: effect of high luminal Ca2+ and phosphorylation.
abstract: Calsequestrin, the major calcium sequestering protein in the sarcoplasmic reticulum of muscle, forms a quaternary complex with the ryanodine receptor calcium release channel and the intrinsic membrane proteins triadin and junctin. We have investigated the possibility that calsequestrin is a luminal calcium concentration sensor for the ryanodine receptor. We measured the luminal calcium concentration at which calsequestrin dissociates from the ryanodine receptor and the effect of calsequestrin on the response of the ryanodine receptor to changes in luminal calcium. We provide electrophysiological and biochemical evidence that: 1), luminal calcium concentration of >/=4 mM dissociates calsequestrin from junctional face membrane, whereas in the range of 1-3 mM calsequestrin remains attached; 2), the association with calsequestrin inhibits ryanodine receptor activity, but amplifies its response to changes in luminal calcium concentration; and 3), under physiological calcium conditions (1 mM), phosphorylation of calsequestrin does not alter its ability to inhibit native ryanodine receptor activity when the anchoring proteins triadin and junctin are present. These data suggest that the quaternary complex is intact in vivo, and provides further evidence that calsequestrin is involved in the sarcoplasmic reticulum calcium signaling pathway and has a role as a luminal calcium sensor for the ryanodine receptor.
authors.affiliation: John Curtin School of Medical Research, Australian Capital Territory, Australia. nicole.beard@anu.edu.au
authors.forename: Nicole A
authors.initials: NA
authors.lastname: Beard
authors.forename: Marco G
authors.initials: MG
authors.lastname: Casarotto
authors.forename: Lan
authors.initials: L
authors.lastname: Wei
authors.forename: Magdolna
authors.initials: M
authors.lastname: Varsányi
authors.forename: Derek R
authors.initials: DR
authors.lastname: Laver
authors.forename: Angela F
authors.initials: AF
authors.lastname: Dulhunty
mesh_terms: D000135:Acid Phosphatase / Q000737:chemistry; D000818:Animals; D002118:Calcium / Q000737:chemistry / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000737:chemistry; D002155:Calsequestrin / Q000737:chemistry* / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry; D047390:Casein Kinase II / Q000737:chemistry; D002845:Chromatography; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004594:Electrophysiology; D005978:Glutathione / Q000378:metabolism; D015151:Immunoblotting; D008051:Lipid Bilayers; D009682:Magnetic Resonance Spectroscopy; D008565:Membrane Proteins / Q000737:chemistry; D006899:Mixed Function Oxygenases / Q000737:chemistry; D009124:Muscle Proteins / Q000737:chemistry; D018482:Muscle, Skeletal / Q000378:metabolism; D009132:Muscles / Q000378:metabolism; D010766:Phosphorylation; D011487:Protein Conformation; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D019837:Ryanodine Receptor Calcium Release Channel / Q000096:biosynthesis* / Q000737:chemistry; D012519:Sarcoplasmic Reticulum / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D002135:Calcium-Binding Proteins; D002155:Calsequestrin; D002352:Carrier Proteins; D008051:Lipid Bilayers; D008565:Membrane Proteins; D009124:Muscle Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D019837:Ryanodine Receptor Calcium Release Channel; C498269:TRDN protein, human; C070048:triadin; C498842:Junctin protein, Oryctolagus cuniculus; D006899:Mixed Function Oxygenases; D047390:Casein Kinase II; D000135:Acid Phosphatase; D005978:Glutathione; D002118:Calcium
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