Steroid hormone metabolizing P-450 in human prostate
| Project/Area Number |
11671575
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Urology
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| Research Institution | Osaka City University Medical School |
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Principal Investigator |
KAWASHIMA Hidenori Osaka City University Medical School, Department of Urology, Research Associate, 医学部, 助手 (70234060)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2000: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1999: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | CYP3A5 / human prostate / unique 5'-untranslated sequence / protein expression / purification / 6β-hydroxylation / testosterone / progesterone / unique 5'-untranslated sequence / P450・3A5 / uniqne 5'-untronslated sequence |
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| Research Abstract |
A cDNA clone coding for CYP3A5 has been isolated from a human prostate cDNA library. The human prostate CYP3A5 cDNA possessed a unique 5'-untranslated sequence, suggesting the prostate specific regulation of CYP3A5 different from liver. Hybridization screening of a human genomic BAC library yielded four positive clones, two of which were shown to contain the unique 5'-untranslated sequence by Southern blot. The CYP3A5 recombinant protein expressed in Escherichia coli using the pCWOri expression vector was purified to an almost electrophoretically homogeneity with a specific content of 4.4 nmol of P450/mg of protein. This purified P450 protein exhibited 6β-hydroxylation activity toward both testosterone and progesterone. No polar metabolite of 5α-dihydrotestosterone (DHT) formed by this P450 was detected. The K_m values for testosterone and progesterone 6β-hydroxylation were 143 and 114μM, respectively, with V_<max> values of 0.48 and 0.21 nmol/min/nmol of P450, respectively. This is the first report that the cDNA for a particular form of P450, CYP3A5, has been isolated from human prostate and that the purified recombinant protein of CYP3A5 has been demonstrated to be active in the metabolism of sex hormones.
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Report
(3 results)
Research Products
(4 results)
