Regulatory differences between Ca(2+)-ATPase in plasma membranes from chicken (nucleated) and pig (anucleated) erythrocytes.
Kinetic and regulatory properties of the plasma membrane Ca(2+)-ATPase activity from chicken (nucleated) erythrocytes were studied and compared to those from pig (anucleated) erythrocytes. In the absence of known activators: (1) Ca(2+) affinity for the Ca(2+)-ATPase activity from nucleated erythrocytes was 12-fold higher than that from pig erythrocytes, and thus the enzyme is sensitive to physiological Ca(2+) concentrations; (2) the enzyme from chicken erythrocytes showed two apparent Km values for ATP, as compared to one apparent Km value displayed by pig erythrocytes; (3) Ca(2+)-ATPase inserted in chicken erythrocyte membranes showed a low sensitivity to activation by phosphatidylinositol-4-phosphate; (4) when p-NPP was used as substrate, the activity of chicken erythrocytes was high, similar to that attained by pig erythrocytes, but barely sensitive to activation by dimethylsulfoxide and calmodulin. ATP hydrolysis was 10-fold lower than that displayed by pig erythrocytes and the maximal velocity was activated three-fold by calmodulin. The enzyme was insensitive to alkaline phosphatase treatment and showed a single phosphorylation band in electrophoresis, ruling out the possibility of previous modulation by endogenous kinases and/or by partial proteolysis. The differences may be attributed to some endogenous modulator, to distinct isoforms, or to a difference in the E(1)/E(2) states of the enzyme.
Alves-Ferreira
Marcelo
M
Departamento de Bioquímica Médica, ICB/CCS, UFRJ, CEP 21941-590, Rio de Janeiro, RJ, Brazil.
da Silva
Elias C C
EC
Ferreira-Pereira
Antonio
A
Scofano
Helena M
HM
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000097:blood* / Q000737:chemistry; D002147:Calmodulin / Q000494:pharmacology; D002645:Chickens / Q000097:blood*; D004789:Enzyme Activation; D004910:Erythrocyte Membrane / Q000201:enzymology*; D013552:Swine / Q000097:blood*
D002147:Calmodulin; D000255:Adenosine Triphosphate; D000252:Calcium-Transporting ATPases; D002118:Calcium