Calsequestrin Is an Inhibitor of Skeletal Muscle Ryanodine Receptor Calcium Release Channels
Beard, Nicole; Sakowska, Magdalena; Dulhunty, Angela; Laver, Derek Rowland
Description
We provide novel evidence that the sarcoplasmic reticulum calcium binding protein, calsequestrin, inhibits native ryanodine receptor calcium release channel activity. Calsequestrin dissociation from junctional face membrane was achieved by increasing luminal (trans) ionic strength from 250 to 500 mM with CsCl or by exposing the luminal side of ryanodine receptors to high [Ca2+] (13 mM) and dissociation was confirmed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western...[Show more]
dc.contributor.author | Beard, Nicole | |
---|---|---|
dc.contributor.author | Sakowska, Magdalena | |
dc.contributor.author | Dulhunty, Angela | |
dc.contributor.author | Laver, Derek Rowland | |
dc.date.accessioned | 2015-12-13T23:26:58Z | |
dc.identifier.issn | 0006-3495 | |
dc.identifier.uri | http://hdl.handle.net/1885/93095 | |
dc.description.abstract | We provide novel evidence that the sarcoplasmic reticulum calcium binding protein, calsequestrin, inhibits native ryanodine receptor calcium release channel activity. Calsequestrin dissociation from junctional face membrane was achieved by increasing luminal (trans) ionic strength from 250 to 500 mM with CsCl or by exposing the luminal side of ryanodine receptors to high [Ca2+] (13 mM) and dissociation was confirmed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. Calsequestrin dissociation caused a 10-fold increase in the duration of ryanodine receptor channel opening in lipid bilayers. Adding calsequestrin back to the luminal side of the channel after dissociation reversed this increased activity. In addition, an anticalsequestrin antibody added to the luminal solution reduced ryanodine receptor activity before, but not after, calsequestrin dissociation. A population of ryanodine receptors ("35%) may have initially lacked calsequestrin, because their activity was high and was unaffected by increasing ionic strength or by anticalsequestrin antibody: their activity fell when purified calsequestrin was added and they then responded to antibody. In contrast to native ryanodine receptors, purified channels, depleted of triadin and calsequestrin, were not inhibited by calsequestrin. We suggest that calsequestrin reduces ryanodine receptor activity by binding to a coprotein, possibly to the luminal domain of triadin. | |
dc.publisher | Biophysical Society | |
dc.source | Biophysical Journal | |
dc.subject | Keywords: binding protein; calsequestrin; protein antibody; ryanodine receptor; article; channel gating; controlled study; dissociation; ionic strength; lipid bilayer; nonhuman; polyacrylamide gel electrophoresis; protein domain; protein protein interaction; sarcop | |
dc.title | Calsequestrin Is an Inhibitor of Skeletal Muscle Ryanodine Receptor Calcium Release Channels | |
dc.type | Journal article | |
local.description.notes | Imported from ARIES | |
local.description.refereed | Yes | |
local.identifier.citationvolume | 82 | |
dc.date.issued | 2002 | |
local.identifier.absfor | 060104 - Cell Metabolism | |
local.identifier.ariespublication | MigratedxPub26426 | |
local.type.status | Published Version | |
local.contributor.affiliation | Beard, Nicole, College of Medicine, Biology and Environment, ANU | |
local.contributor.affiliation | Sakowska, Magdalena, College of Medicine, Biology and Environment, ANU | |
local.contributor.affiliation | Dulhunty, Angela, College of Medicine, Biology and Environment, ANU | |
local.contributor.affiliation | Laver, Derek Rowland, College of Medicine, Biology and Environment, ANU | |
local.description.embargo | 2037-12-31 | |
local.bibliographicCitation.startpage | 310 | |
local.bibliographicCitation.lastpage | 320 | |
dc.date.updated | 2015-12-12T09:48:33Z | |
local.identifier.scopusID | 2-s2.0-0036219539 | |
Collections | ANU Research Publications |
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