PHYSIOLOGICAL ROLE OF HUMAN CYTOCHROME P450 FAMILY 4
| Project/Area Number |
06680630
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Functional biochemistry
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| Research Institution | Fukuyma University |
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Principal Investigator |
KUSUNOSE Masamichi Fukuyama University, Faculty of engineering, Professor, 工学部, 教授 (10046766)
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| Co-Investigator(Kenkyū-buntansha) |
KIKUTA Yasushi Fukuyama University, Faculty of engineering, Associate, 工学部, 助手 (50224895)
YAMAMOTO Satoru Fukuyama University, Faculty of engineering, Associate Professor, 工学部, 助教授 (10191404)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1995: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1994: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | omega-Hydroxylation / Cytochrome P450 / Leukotriene B_4 / Fatty acids / Leukocytes / Gene / Liver / ロイコトリエン |
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| Research Abstract |
1. We have purified to homogeneity a fatty acid omega-hydroxylase (cytochrome P450HLomega) from human liver microsomes. We have also isolated a cDNA for human liver fatty acid omega-hydroxylase (P4504A11) using the cDNA for rabbit fatty acid omega-hydroxylase (P4504A7) as a probe. This cDNA encoded a protein of 519 amino acids with a molecular weight of 59,347. The protein expressed in yeast cells transfected with the cDNA catalyzed the omega-hydroxylation of vairous fatty acids. 2. We have found for the first time that human liver microsomes are highly active in the omega-hydroxylation of leukotriene B4 (LTB4). We have isolated and sequenced a cDNA for LTB4 omega-hydroxylase using the cDNA for human neutrophil LTB4 omega-hydroxylase (P4504F3) as a probe from a human liver cDNA library. The cDNA-deduced amino acid sequence showed 87.3% homology to that of human neutrophil LTB4 omega-hydroxylase (P4504F3), and this human liver P450 was designated P4504F2. The solubilized microsomes from yeast cells transfected with the cDNA rapidly catalyzed the omega-hydroxylation of 6-trans LTB4 and lipoxin A4 as well as LTB4.3. Human neutrophil LTB4 omega-hydroxylase (P4504F3) was purified to an electrophoretically homogenous state from the microsomes of yeast cells transfected with the cDNA by chromatography on AH-Sepharose 4B,DEAE-5pw HPLC and hydroxylapatite HPLC columns. The purified enzyme catalyzed the omega-hyroxylation of LTB4,6-trans LTB4 and lipoxin B4.4. FISH analysis indicated that P4504F3 gene lies on human chromosome 19p13.2.
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Report
(3 results)
Research Products
(6 results)
