Two major antigens of heart sarcolemma are Ca2(+)-binding glycoproteins that copurify with the dihydropyridine receptor.
Ca2+ binding has been studied in isolated heart sarcolemmal membranes using the 45Ca overlay technique. 45Ca bound to two sarcolemmal polypeptides of 125 kDa and 97 kDa in preparations from dog, rabbit, cow and pig. During fractionation on DEAE ion-exchange and wheat-germ lectin affinity columns, the two Ca2(+)-binding polypeptides copurified with the dihydropyridine receptor associated with the voltage gated Ca2+ channel. These polypeptides were the major proteins in the isolated fraction as judged by silver staining in SDS-PAGE. Antisera raised against purified dog heart, sarcolemma indicated that the 125 and 97 kDa polypeptides were highly antigenic components of this membrane. The antisera cross-reacted with similar polypeptides in cardiac sarcolemmal preparations from rabbit, cow and pig, but not sarcoplasmic reticulum membranes. Purified antibodies against the 125 kDa polypeptide did not cross-react with the 97 kDa polypeptide, while antibodies against the 97 kDa polypeptide did not cross-react with the 125 kDa polypeptide. Both the 125 kDa and 97 kDa polypeptides bound wheat-germ lectin, suggesting both were glycoproteins. It is unlikely that these Ca2+ binding glycoproteins represent subunits of the dihydropyridine receptor-Ca2+ channel in this membrane.
Tuana
B S
BS
Department of Pharmacology, University of Ottawa, Canada.
Sunahara
R K
RK
Murphy
B J
BJ
Pierce
G N
GN
D000818:Animals; D000941:Antigens / Q000302:isolation & purification*; D015220:Calcium Channels; D002132:Calcium Radioisotopes; D002135:Calcium-Binding Proteins / Q000302:isolation & purification*; D002417:Cattle; D003429:Cross Reactions; D004285:Dogs; D006023:Glycoproteins / Q000302:isolation & purification*; D007106:Immune Sera / Q000096:biosynthesis; D015151:Immunoblotting; D009206:Myocardium / Q000276:immunology*; D011817:Rabbits; D011978:Receptors, Nicotinic / Q000302:isolation & purification*; D012508:Sarcolemma / Q000276:immunology*; D012519:Sarcoplasmic Reticulum / Q000276:immunology; D013552:Swine
D000941:Antigens; D015220:Calcium Channels; D002132:Calcium Radioisotopes; D002135:Calcium-Binding Proteins; D006023:Glycoproteins; D007106:Immune Sera; D011978:Receptors, Nicotinic