2006 Fiscal Year Final Research Report Summary
Identification and functional analysis of two unknown PHGPx in embryogenesis
| Project/Area Number |
17590067
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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 |
Biological pharmacy
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| Research Institution | Kitasato University |
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Principal Investigator |
IMAI Hirotaka Kitasato University, School of Pharmaceutical Sciences, Associate Professor, 薬学部, 助教授 (50255361)
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| Project Period (FY) |
2005 – 2006
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| Keywords | Embryogenesis / Oxidative stress / Transgenic rescue / Proteome Analysis / Lipid / Glutathione Peroxidase |
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| Research Abstract |
Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is a unique antioxidant enzyme that can directly reduce phospholipid hydroperoxide. Disruption of PHGPx gene in mice resulted in the embryonic lethality at 8.5 dpc. However, the analysis of role of PHGPx in embryogenesis has remained.
In this study, to clarify the role of PHGPx during embryogenesis, we examined about the expression of PHGPx protein, mRNA in embryogenesis and the functional analysis using in vitro culture system and transgenic rescue method.
We found that new 18kDa PHGPx was expressed in 7.5dpc-9.5dpc embryos.
This PHGPx was associated with other protein via disulfide formation.
From 5'RACE analysis, new transcriptional star site in lb exons was found and new translational start site. In vitro embryo culture system, PHGPx KO embryo died and could not formation of ICM, however transfection of non-mitochondrial PHGPx rescued lethality of KO embryo. We developed new strategy for analysis of role of PHGPx during embryogenesis using transgenic rescue. Trans gene of normal PHGPx gene rescued lethality of KO mice, but all mutant of PHGPx gene did not rescued lethality of KO mice. In placenta we found that 20kDa PHGPx was associated with 10kDa other protein and dimmer formation.
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