Localization and function of enzymes: role of arginine kinase in cilia of Tetrahymena pyriformis
Project/Area Number |
24570087
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Multi-year Fund |
Section | 一般 |
Research Field |
Animal physiology/Animal behavior
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Research Institution | Kochi University |
Principal Investigator |
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Project Period (FY) |
2012-04-01 – 2015-03-31
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Project Status |
Completed (Fiscal Year 2015)
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Budget Amount *help |
¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000)
Fiscal Year 2015: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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Keywords | 酵素の局在 / アルギニンキナーゼ / 繊毛運動 / ミリストイル化 / アルギニンリン酸シャトル / 無細胞タンパク質合成 / 酵素機能 |
Outline of Final Research Achievements |
Tetrahymena pyriformis contains two arginine kinases (AKs), a typical 40-kDa AK (AK1) and a two-domain 80-kDa enzyme (AK2). Interestingly, the amino acid sequence of T. pyriformis AK1, had a distinct myristoylation signal sequence at the N-terminus. In this work, we isolated and cloned the N-myristoyltransferase (NMT) cDNA from T. pyriformis. Then the NMT assay were performed, using the recombinant NMT and the N-terminal peptide GCSNSRSG of T. pyriformis AK1 or the peptide CSNSRSG which lacks N-terminal G of the former peptide. The use of the former peptide gave a typical Michaelis-Menten curve, while that for the latter gave no activity. A sufficient amount of modified AK1 enzyme was synthesized using an insect cell-free protein synthesis system. Then peptide mass fingerprinting (PMF) analyses were performed. The target N-myristoylated peptide was clearly observed with an experimental mass (m/z-H+) of 832.4747, indicating that the synthesized T. pyriformis AK1 is myristoylated.
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Report
(4 results)
Research Products
(10 results)
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[Journal Article] Two arginine kinases of Tetrahymena pyriformis: characterization and localization.2014
Author(s)
Michibata, J., Okazaki, N., Motomura, S., Uda, K., Fujiwara, S. and Suzuki, T.
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Journal Title
Comp. Biochem. Physiol. Part B.
Volume: 171
Pages: 34-41
DOI
Related Report
Peer Reviewed / Acknowledgement Compliant
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