Title of article :
cDNA Cloning, Heterologous Expression, and Characterization of Mouse CYP2G1, an Olfactory-Specific Steroid Hydroxylase
Author/Authors :
Hua، نويسنده , , Zichun and Zhang، نويسنده , , Qing-yu and Su، نويسنده , , Ting and Lipinskas، نويسنده , , Thomas W. and Ding، نويسنده , , Xinxin، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 1997
Pages :
7
From page :
208
To page :
214
Abstract :
CYP2G1 is expressed specifically in the olfactory mucosa in rabbits and rats. In the present study, a full-length cDNA for mouse CYP2G1 was obtained using a PCR approach with RNA preparations from the olfactory mucosa of C57BL/6 mice. Sequence comparisons indicated that mouse CYP2G1 is highly homologous in deduced amino acid sequence to rabbit (82.4% identity) and rat CYP2G1 (94.9% identity). RNA blot and immunoblot analyses indicated that mouse CYP2G1 is expressed only in the olfactory mucosa. The coding region of the mouse CYP2G1 cDNA was cloned into a baculoviral expression vector for heterologous production of the enzyme in cultured insect cells. Heterologously expressed mouse CYP2G1 was active in a reconstituted system toward testosterone and progesterone, producing all the major metabolites detected in olfactory microsomal reactions, including 15α-, 15β-, and 2β-hydroxytestosterone from testosterone and two unidentified metabolites from progesterone. Kinetic analysis indicated that mouse CYP2G1 has relatively high affinities toward the steroid substrates, withKmvalues in the micromolar range for both testosterone and progesterone. At a substrate concentration of 10 μm, microsomes of olfactory mucosa had much higher turnover numbers toward testosterone and progesterone than hepatic microsomes, consistent with the olfactory-specific expression of a high-affinity sex steroid hydroxylase. These findings will facilitate further molecular genetics studies on the biological function of CYP2G1 in a mouse model.
Keywords :
heterologous expression , CYP2G1 , Steroid hormone , steroid metabolism , Olfactory mucosa , mouse , cDNA cloning
Journal title :
Archives of Biochemistry and Biophysics
Serial Year :
1997
Journal title :
Archives of Biochemistry and Biophysics
Record number :
1608765
Link To Document :
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