Olfactory adenylyl cyclase. Identification and purification of a novel enzyme form.
Rat olfactory adenylyl cyclase has been identified by means of a monoclonal antibody BBC-2, which reacts with both Ca2+/calmodulin-sensitive and -insensitive forms of adenylyl cyclase (Mollner, S., and Pfeuffer, T. (1988) Eur. J. Biochem. 171, 265-271). The antibody recognized a 180-kDa polypeptide in olfactory cilia but not in decilitated olfactory epithelial membranes. A protein of the same mobility was observed when olfactory adenylyl cyclase was purified by forskolin-agarose affinity chromatography followed by radioiodination. Its identity was further established by cross-linking to [32P]ADP-ribosylated Gs alpha (GTP-binding protein), to yield a single radiolabeled product of Mr approximately 220. Olfactory adenylyl cyclase has a approximately 3-fold higher turnover number, as assessed from stoichiometric binding of [35S]guanosine 5'-(3-O-thio)triphosphate. Therefore, the considerably higher specific adenylyl cyclase activity in olfactory cilia must be due to a approximately 100-fold higher molar concentration of enzyme in this tissue.
Pfeuffer
E
E
Department of Physiological Chemistry, University of Würzburg, School of Medicine, Federal Republic of Germany.
Mollner
S
S
Lancet
D
D
Pfeuffer
T
T
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000262:Adenylyl Cyclases / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology; D002118:Calcium / Q000494:pharmacology; D002147:Calmodulin / Q000494:pharmacology; D002846:Chromatography, Affinity; D002923:Cilia / Q000201:enzymology*; D005576:Colforsin / Q000494:pharmacology; D004848:Epithelium / Q000201:enzymology; D019204:GTP-Binding Proteins / Q000378:metabolism; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate / Q000378:metabolism / Q000494:pharmacology; D008970:Molecular Weight; D009831:Olfactory Mucosa / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains; D013873:Thionucleotides / Q000378:metabolism / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D002147:Calmodulin; D013873:Thionucleotides; D005576:Colforsin; D000246:Adenosine Diphosphate Ribose; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases; D002118:Calcium