title: Peptide sequence analysis and molecular cloning reveal two calcium pump isoforms in the human erythrocyte membrane.
abstract: The sequence of more than 1,000 amino acid residues, derived from two different isoforms, has been determined from peptides generated from purified human erythrocyte membrane Ca2(+)-ATPase (hPMCA). Several of these peptide sequences correspond to the previously reported, cDNA deduced sequence of the "teratoma" Ca2+ pump isoform hPMCA1 (Verma, A. K., Filoteo, A. G., Stanford, D. R., Wieben, E. D., Penniston, J. T., Strehler, E. E., Fischer, R., Heim, R., Vogel, G., Matthews, S., Strehler-Page, M.-A., James, P., Vorherr, T., Krebs, J., and Carafoli, E. (1988) J. Biol. Chem. 263, 14152-14159). The complete primary structure of a novel isoform (hPMCA3) has been determined by molecular cloning and nucleotide sequencing of its corresponding cDNA. This new member of the plasma membrane Ca2+ pump family consists of 1,205 amino acid residues with a calculated Mr of 133,930, and it shows 88% similarity (75% identity) with the previously sequenced pump isoform. Specific probes detect major mRNA species of 5.6 kilobases for hPMCA1, and of 7.5 kilobases for hPMCA3, on Northern blots of human K562 erythroleukemic cell RNA. A large number of peptide sequences match perfectly with only one or the other of these isoforms and all peptides (with 6 exceptions corresponding to a contaminant protein or to a third minor Ca2+ pump isoform) are found in either only one or in both of the isoforms. The two erythrocyte Ca2+ pumps display high sequence divergence in a few localized regions that may determine isoform-specific functional specializations; for example, the putative extracellular loop separating transmembrane domains 1 and 2, the highly negatively charged region previously suggested to be involved in Ca2+ binding, and the site of cAMP-dependent protein kinase phosphorylation.
authors: Laboratory for Biochemistry, Swiss Federal Institute of Technology, Zurich.
authors: E E
authors: EE
authors: Strehler
authors: P
authors: P
authors: James
authors: R
authors: R
authors: Fischer
authors: R
authors: R
authors: Heim
authors: T
authors: T
authors: Vorherr
authors: A G
authors: AG
authors: Filoteo
authors: J T
authors: JT
authors: Penniston
authors: E
authors: E
authors: Carafoli
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D000252:Calcium-Transporting ATPases / Q000097:blood / Q000235:genetics*; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D004910:Erythrocyte Membrane / Q000201:enzymology*; D015723:Gene Library; D006801:Humans; D007413:Intestinal Mucosa / Q000201:enzymology; D007527:Isoenzymes / Q000097:blood / Q000235:genetics*; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid
chemical_list: D007527:Isoenzymes; D004247:DNA; D000252:Calcium-Transporting ATPases