Evidence that platelet and skeletal sarcoplasmic reticulum Ca2+-ATPase are structurally distinct.
Proteolytic digestion and indirect immunostaining were used to compare the platelet and sarcoplasmic reticulum Ca2+-ATPase proteins. When the platelet and sarcoplasmic reticulum Ca2+-ATPase proteins were digested in the native state with trypsin, the platelet Ca2+-ATPase, which had an apparent undigested molecular mass of 103 kDa, yielded 78-kDa and 25-kDa fragments. Calcium transport activity depended on the integrity of the 103-kDa protein, while the digested protein had residual ATPase activity. Tryptic digestion of the sarcoplasmic reticulum pump protein, which also had an undigested molecular mass of 103 kDa, yielded products with apparent molecular masses of 55 kDa, 36 kDa, and 26 kDa. Distinct patterns were also observed when the platelet and sarcoplasmic reticulum calcium pump proteins were digested with chymotrypsin and Staphylococcus aureus protease in the presence of sodium dodecyl sulfate. Chymotrypsin digestion of the platelet protein resulted in the appearance of products with apparent molecular masses of 70 kDa, 39 kDa, and 31 kDa, while a similar digestion of the sarcoplasmic reticulum calcium pump protein yielded 54-kDa, 52.5-kDa, 46-kDa, 41-kDa, and 36-kDa fragments. Exposure of the sarcoplasmic reticulum and platelet Ca2+-ATPase proteins to S. aureus protease also yielded dissimilar fragmentation patterns. These results indicate that the Ca2+-ATPases from platelets and sarcoplasmic reticulum are distinct proteins.
Fischer
T H
TH
Campbell
K P
KP
White
G C
GC
D000818:Animals; D001792:Blood Platelets / Q000201:enzymology*; D002118:Calcium / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000032:analysis* / Q000502:physiology; D006801:Humans; D008970:Molecular Weight; D011487:Protein Conformation; D051381:Rats; D012519:Sarcoplasmic Reticulum / Q000201:enzymology*; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D012995:Solubility; D014357:Trypsin / Q000494:pharmacology
D012967:Sodium Dodecyl Sulfate; D014357:Trypsin; D000252:Calcium-Transporting ATPases; D002118:Calcium