Cloning and expression of a cDNA coding for a rat liver plasma membrane ecto-ATPase. The primary structure of the ecto-ATPase is similar to that of the human biliary glycoprotein I.
The amino acid sequence of the ecto-ATPase from rat liver was deduced from analysis of cDNA clones and a genomic clone. Immunoblots with antibodies raised against a peptide sequence deduced from the cDNA sequence indicated that the determined amino acid sequence is that of the ecto-ATPase. The deduced sequence predicts a 519-amino acid protein with a calculated molecular mass of 57,388 daltons. There are 16 potential asparagine-linked glycosylation sites in the protein. Hydropathy analysis of the deduced amino acid sequence indicates that the protein has two hydrophobic stretches. One is located at the N-terminal and the other is near the C-terminal end. A full-length clone encoding the ecto-ATPase was expressed transiently in mouse L cells and human HeLa cells. The cell lysate from the transfected cells contained immunoreactive ecto-ATPase and Ca2+-stimulated ATPase activities. The expressed protein is glycosylated and has an apparent molecular weight (100,000) similar to that of the rat liver plasma membrane ecto-ATPase.
Lin
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Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, Massachusetts 02138.
Guidotti
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D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD; D001483:Base Sequence; D000252:Calcium-Transporting ATPases / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D015815:Cell Adhesion Molecules; D002462:Cell Membrane / Q000201:enzymology; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification*; D006023:Glycoproteins; D006367:HeLa Cells; D006801:Humans; D007106:Immune Sera; D007739:L Cells; D008099:Liver / Q000201:enzymology*; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D051381:Rats; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection
D015703:Antigens, CD; C069847:CD66 antigens; D015815:Cell Adhesion Molecules; D006023:Glycoproteins; D007106:Immune Sera; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D004247:DNA; D000252:Calcium-Transporting ATPases