Molecular cloning and expression of a cDNA encoding an olfactory-specific mouse phenol sulphotransferase.
Previously we demonstrated the presence of phenol sulphotransferase (P-ST) in mouse nasal cytosols and identified its zonal location in mouse nasal cavity by staining with an antiserum raised against a rat liver P-ST isoenzyme, PSTg. In the present study a cDNA was isolated from a mouse olfactory cDNA library by immunological screening with the antiserum. The isolated cDNA consisted of 1347 bp with a 912 bp open reading frame encoding a 304-residue polypeptide. Both the nucleotide and deduced amino acid sequences of the cDNA were 94% identical with those of a rat liver P-ST isoenzyme, ST1C1. The expressed enzyme in Escherichia coli displayed high P-ST activity towards phenolic odorants such as eugenol and guaiacol, and it showed a high N-hydroxy-2-acetylaminofluorene sulphation activity in comparison with the rat ST1C1 enzyme. These results indicate that the olfactory P-ST encoded by the cDNA is a mouse orthologue of rat ST1C1; however, expression of the olfactory P-ST mRNA is specific for nasal tissues as revealed by reverse transcriptase-mediated PCR (RT-PCR).
Tamura
H O
HO
Kyoritsu College of Pharmacy, 1-5-30, Shibakoen, Minato-ku, Tokyo 105, Japan.
Harada
Y
Y
Miyawaki
A
A
Mikoshiba
K
K
Matsui
M
M
D000595:Amino Acid Sequence; D000818:Animals; D015239:Arylsulfotransferase / Q000737:chemistry*; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005054:Eugenol / Q000378:metabolism; D006139:Guaiacol / Q000378:metabolism; D006881:Hydroxyacetylaminofluorene / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D009297:Nasal Mucosa / Q000201:enzymology*; D010802:Phylogeny; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
D012333:RNA, Messenger; D011994:Recombinant Proteins; D005054:Eugenol; D006881:Hydroxyacetylaminofluorene; D006139:Guaiacol; D015239:Arylsulfotransferase