Characterization of soluble platelet guanylyl cyclase with peptide antibodies.
Soluble guanylyl cyclase partially purified from bovine and human platelets was characterized with antibodies raised against synthetic peptides corresponding to different sequences of the alpha 1- and beta 1-subunits of the bovine lung enzyme. On immunoblots, the platelet guanylyl cyclase was recognized by the four antisera used, with the exception of an antiserum against the C-terminus of the beta 1-subunit which did not react with the human platelet but with the bovine platelet beta 1-subunit. Furthermore the human platelet beta 1-subunit exhibited a slightly lower molecular mass than the bovine protein. The C-terminal antibodies precipitated native platelet and lung guanylyl cyclase activity. In contrast an antibody against a peptide out of the putative catalytic domain, which is highly conserved between all guanylyl cyclases sequenced so far, did not precipitate native guanylyl cyclase, although it recognized both subunits on immunoblots, suggesting that the respective amino acid sequence is located in an inner site of the protein.
Institut für Pharmakologie, Freie Universität Berlin, Federal Republic of Germany.
F
F
Guthmann
B
B
Mayer
D
D
Koesling
W R
WR
Kukovetz
E
E
Böhme
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D001792:Blood Platelets / Q000201:enzymology* / Q000276:immunology; D015153:Blotting, Western; D002417:Cattle; D003600:Cytosol / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006162:Guanylate Cyclase / Q000097:blood* / Q000276:immunology; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology*; D011233:Precipitin Tests
D000906:Antibodies; D010455:Peptides; D006162:Guanylate Cyclase