2002 Fiscal Year Final Research Report Summary
ATP-dependent proteolysis in mitochondria
| Project/Area Number |
13640683
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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 |
動物生理・代謝
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| Research Institution | Yamaguchi University |
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Principal Investigator |
WATABE Shoji Yamaguchi University School of Medicine, Professor, 医学部, 教授 (30113020)
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| Co-Investigator(Kenkyū-buntansha) |
YAMASHIRO Yasuhiro Yamaguchi University School of Medicine, Assistant Professor, 医学部, 講師 (50243671)
YAMAMOTO Yoshimi Yamaguchi University Fuculty of Agriculture, Professor, 農学部, 教授 (40115514)
TAKAHASHI Susumu Yamaguchi University Fuculty of Agriculture, Professor, 農学部, 教授 (90022665)
HASEGAWA Hiroyuki Teikyo University of Science & Technology, Undergraduate School of Science & Engineering, Professor, 理工学部, 教授 (10092983)
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| Project Period (FY) |
2001 – 2002
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| Keywords | mitochondria / protease / gene expression / adrenal cortex / proteolysis / GFP |
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| Research Abstract |
We have investigated about ATP-dependent proteolytic system in mitochondria. Although there are some ATP-dependent proteases in animal mitochondria, we focused our studies on ATP-dependent LON protease. We purified the protease from bovine adrenal cortex and characterized it. In the present research we found that the enzyme was stimulated by peptides or proteins and that the stimulation was due to the stimulation of release of ADP (one of the reaction products) from the enzyme. We determined nucleotide sequence of the cDNA encoding the protease. The open reading frame of the cDNA contained 960 amino acids including presequence. To determine the precise length of the presequence we determined N-terminal amino acid sequence of the mature enzyme. We revealed that the enzyme has heterogeneous N-terminal starting from an amino acid between Trp-69 and Asn-73. To determine whether the presequence contained the information for transporting the protein into mitochondria, we expressed a fusion protein composed of the presequence and Green Fluorescent Protein (GFP) in Chinese hamster ovary cells and found that the green fluorescence was incorporated into granules which were assumed to be mitochondria.
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[Publications] Yamahama, Y., Uto, N., Tamotsu, S., Miyata, T., Yamamoto, Y., Watabe, S., Takahashi, S.Y.: "Invivo activation of pro-form Bombix cysteine proteinase (BCP) in silkworm eggs : localization of yolk proteins and BCP, and acidification of yolk granules"J. Insect Physiol.. 49. 131-140 (2003)
Description
「研究成果報告書概要(和文)」より
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