| Project/Area Number |
10640636
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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 |
植物生理
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
WADA Hajime Graduate School of Science, Kyushu University Associate Professor, 大学院・理学研究科, 助教授 (60167202)
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| Co-Investigator(Kenkyū-buntansha) |
和田 元 九州大学, 大学院・理学研究科, 助教授 (60167202)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 1999: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1998: ¥2,700,000 (Direct Cost: ¥2,700,000)
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| Keywords | Arabidopsis / Fatty acid symthesis / Lipoic acid / Lipoic acid symthase / Lipoyltransferare / ミトコンドリア / シロイヌナズナ / 分子生物学 |
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| Research Abstract |
Lipoic acid is a coenzyme that is essential for activity of enzyme complexes such as the pyruvate dehydrogenase and glycine decarboxylase complexes. Despite the importance of the lipoyl prosthetic group in the function of the enzyme complexes involved in central metabolism, the biosynthesis and transter of lipoic acid to cognate proteins have not been well understood in any organism.
In this study I have isolated and characterized LIP1 and LIP2 cDNAs for lipoic acid synthase and lipoyltransferase, respectively, of Arabidopsis thaliana. The LIP1 cDNA was shown to code for Arabidopsis lipoic acid synthase by its ability to complement an E. coli mutant that is defective in lipoic acid synthase. DNA sequence analysis of the LIP1 cDNA revealed an open-reading frame predicting a protein of 374 amino acids. Southern hybridization analysis suggests that LIP1 is a single0copy gene in Arabidopsis. Western blot analysis with an antibody against lipoic acid synthase demonstrates that lipoic acid synthase is located in the mitochondrial compartment in Arabidopsis cells as a 43 KD polypeptide. A LIP2 cDNA encoding lipoyltransferase has also been cloned from Arabidopdis. This cDNA was shown to code for lipoyltransterase by its ability to complement a mutant of E. coli defective in lipoyltransferases (Lpla and LipB). DNA sequence analysis of the LIP2 cDNA revealed an open-reading frame predicting a protein of 235 amino acids. Southern and northern hybridization analyses suggest that LIp2 is a single-copy gene and expressed as a mRNA of 860 rucleotides in Arabidopsis leaves. Westem blot analysis with an antibody against lipoyltransferase demonstrates that lipoyltransferase is located in the mitochondrial compartment in Arabidopsis cells as a 29 kDa polypeptide.
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