Cloning and expression of isoform 2 of the human plasma membrane Ca2+ ATPase. Functional properties of the enzyme and its splicing products.
Full-length cDNAs for the three human plasma membrane Ca2+ pump isoforms 2 (PMCA2) differently spliced at the A site were constructed and transferred to baculovirus. The corresponding proteins were expressed after infection in Sf9 insect cells. The proteins were expressed at high levels and retained the canonical properties of the plasma membrane Ca2+ pump. The alternative splicing process failed to produce functional differences detectable with the methods used. The Ca(2+)-dependent ATPase activity of the PMCA2 pumps had a 5-10-fold higher affinity for calmodulin than the PMCA4 pump expressed in the same system. Experiments on the formation of the phosphoenzyme intermediate from ATP revealed that the PMCA2 pumps had higher affinity for ATP than did the PMCA4 counterpart. The response of the two pump types to activating acidic phospholipids was the same.
Hilfiker
H
H
Institute of Biochemistry, Swiss Federal Institute of Technology (ETH), Zürich.
Guerini
D
D
Carafoli
E
E
D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D000252:Calcium-Transporting ATPases / Q000235:genetics / Q000502:physiology*; D002147:Calmodulin / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology; D003001:Cloning, Molecular; D004789:Enzyme Activation; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D010743:Phospholipids / Q000494:pharmacology; D010766:Phosphorylation; D018411:Spodoptera
D002147:Calmodulin; D007527:Isoenzymes; D010743:Phospholipids; D000255:Adenosine Triphosphate; D000252:Calcium-Transporting ATPases