Monoclonal antibody to skeletal muscle ryanodine receptor detects a polypeptide in rat brain: comparison of immunogenic fragments after limited proteolysis.
A polypeptide of high molecular mass has been detected in mammalian brain by a monoclonal antibody, 5C3, raised against skeletal muscle ryanodine receptor. 5C3 does not crossreact with the cardiac ryanodine receptor, the isoform which is believed to be located in many regions of the brain. Endogenous proteases in brain formed a prominent immunogenic fragment of 116 kDa whereas five immunostaining polypeptides greater than 200 kDa were observed in skeletal muscle. Mild trypsin digestion of brain microsomes resulted in fragments of approximately 400 and approximately 280 kDa, of similar mass to two peptides formed from the skeletal muscle ryanodine receptor. However a peptide of 28 kDa, resistant to trypsin, was observed in muscle but not in brain. The brain polypeptide recognised by 5C3 is therefore not identical to the skeletal muscle ryanodine receptor.
Division of Neuroscience, John Curtin School of Medical Research, Australian National University, Canberra.
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Junankar
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Dulhunty
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D015153:Blotting, Western; D001923:Brain Chemistry; D015220:Calcium Channels / Q000032:analysis* / Q000276:immunology; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D008970:Molecular Weight; D009124:Muscle Proteins / Q000032:analysis* / Q000276:immunology; D009132:Muscles / Q000737:chemistry*; D010455:Peptides / Q000032:analysis* / Q000276:immunology; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D019837:Ryanodine Receptor Calcium Release Channel; D014357:Trypsin / Q000378:metabolism
D000911:Antibodies, Monoclonal; D015220:Calcium Channels; D009124:Muscle Proteins; D010455:Peptides; D019837:Ryanodine Receptor Calcium Release Channel; D014357:Trypsin