Linked Life Data

8385488

Source:http://linkedlifedata.com/resource/pubmed/id/8385488

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
14
pubmed:dateCreated
1993-5-13
pubmed:abstractText
A full-length cDNA encoding the ryanodine receptor of rabbit skeletal muscle sarcoplasmic reticulum was transiently expressed in COS-1 cells. Immunoblotting studies showed that the expressed ryanodine receptor and the native ryanodine receptor of rabbit skeletal muscle were indistinguishable in molecular size and immunoreactivity. Scatchard analysis of [3H]ryanodine binding to transfected COS-1 cell microsomes resulted in a Bmax of 0.22 pmol/mg of protein and a Kd of 16.2 nM. Expressed ryanodine receptors were solubilized in CHAPS and were shown to cosediment with native ryanodine receptors in a sucrose density gradient. Thus, the expressed receptor, like the native receptor, is assembled as a large oligomeric complex. Single-channel recordings in planar lipid bilayers were used to investigate the functional properties of the sucrose gradient-purified complex. The expressed ryanodine receptor formed a large conductance channel activated by ATP and Ca2+ and inhibited by Mg2+ and ruthenium red. Ryanodine reduced the conductance and increased the mean open time in a manner consistent with that of native channels. These results demonstrated that functional binding sites for the physiological ligands (Ca2+, Mg2+, and ATP) and pharmacological ligands (ruthenium red and ryanodine) controlling gating of the Ca2+ release channel are encoded in the ryanodine receptor cDNA and are faithfully expressed in COS-1 cells. Ryanodine receptors expressed in COS-1 cells displayed several conductance states > or = 1 nS not present in native channels. Such anomalous conductance states of the expressed channel might be referable to lack of muscle-specific posttranslational processing or to the need for components not present in COS-1 cells, which may be required to stabilize the channel structure.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
13
pubmed:volume
32
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
3743-53
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:8385488-Adenosine Triphosphate, pubmed-meshheading:8385488-Animals, pubmed-meshheading:8385488-Calcium Channels, pubmed-meshheading:8385488-Cell Line, pubmed-meshheading:8385488-Centrifugation, Density Gradient, pubmed-meshheading:8385488-Cholic Acids, pubmed-meshheading:8385488-DNA, pubmed-meshheading:8385488-DNA Restriction Enzymes, pubmed-meshheading:8385488-Electric Conductivity, pubmed-meshheading:8385488-Gene Expression, pubmed-meshheading:8385488-Immunohistochemistry, pubmed-meshheading:8385488-Muscle Proteins, pubmed-meshheading:8385488-Muscles, pubmed-meshheading:8385488-Plasmids, pubmed-meshheading:8385488-Rabbits, pubmed-meshheading:8385488-Ryanodine, pubmed-meshheading:8385488-Ryanodine Receptor Calcium Release Channel, pubmed-meshheading:8385488-Sarcoplasmic Reticulum, pubmed-meshheading:8385488-Solubility, pubmed-meshheading:8385488-Transfection
pubmed:year
1993
pubmed:articleTitle
Functional expression of cDNA encoding the Ca2+ release channel (ryanodine receptor) of rabbit skeletal muscle sarcoplasmic reticulum in COS-1 cells.
pubmed:affiliation
Banting and Best Department of Medical Research, University of Toronto, C.H. Best Institute, Ontario, Canada.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't