Identification and biochemical analysis of novel olfactory-specific cytochrome P-450IIA and UDP-glucuronosyl transferase.
Two major transmembranal polypeptides of bovine olfactory epithelium were identified by SDS electrophoretic analysis of Triton X-114 solubilized membranes. Both polypeptides were present in large amounts in membranes of the olfactory epithelium but were barely detectable in membranes of the nasal respiratory epithelium. Both polypeptides are enriched in the deciliated epithelium as compared with isolated cilia. One of them is a glycoprotein with an apparent molecular mass of 56 kDa (gp56); the other is an unglycosylated protein with an apparent molecular mass of 52 kDa (p52). Sequence analysis of peptides obtained by CNBr cleavage of purified gp56 indicates that it is highly homologous to UDP-glucuronosyl transferase (UDPGT). Parallel analysis shows that p52 is highly homologous to cytochrome P-450 sequences of the IIA subfamily. This protein is assigned the name P-450olf2. Polyclonal antibodies were raised against synthetic peptides corresponding to gp56 and p52 peptide sequences. Immunoblots with these antibodies reveal the following properties of gp56 and p52: (1) they are enriched in the microsomal fraction of the bovine olfactory epithelium; (2) they are possibly specific to the olfactory epithelium, as we could not detect reactivity in microsomes derived from respiratory epithelium or lung, and only a very small amount of basal reactivity was seen with liver microsomes; (3) cross-reacting proteins exist in microsomes derived from the rat olfactory epithelium. These results are consistent with a mechanism whereby the microsomal enzymes are involved in odorant modification and clearance from the nasal tissue.
Lazard
D
D
Department of Membrane Research, Weizmann Institute of Science, Rehovot, Israel.
Tal
N
N
Rubinstein
M
M
Khen
M
M
Lancet
D
D
Zupko
K
K
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002923:Cilia / Q000201:enzymology; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis*; D004848:Epithelium / Q000201:enzymology; D015698:Genomic Library; D014453:Glucuronosyltransferase / Q000032:analysis*; D008562:Membrane Glycoproteins / Q000032:analysis*; D008969:Molecular Sequence Data; D009296:Nasal Cavity / Q000201:enzymology*; D009297:Nasal Mucosa / Q000201:enzymology; D011817:Rabbits; D051381:Rats; D011919:Rats, Inbred Strains
D008562:Membrane Glycoproteins; D003577:Cytochrome P-450 Enzyme System; D014453:Glucuronosyltransferase