| Project/Area Number |
11680592
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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 |
Bioorganic chemistry
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| Research Institution | Osaka City University |
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Principal Investigator |
IIO Hideo Osaka City University, Graduate School of Science, Professor, 大学院・理学研究科, 教授 (80145771)
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| Co-Investigator(Kenkyū-buntansha) |
HARUMOTO Terue Nara Women's University, Faculty of Science, Associate Professor, 理学部, 助教授 (80198936)
USUKI Yoshinosuke Osaka City University, Graduate School of Science, Lecturer, 大学院・理学研究科, 講師 (30244651)
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| Project Period (FY) |
1999 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2001: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2000: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1999: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | ciliate / Blepharisma / conjugation / glycoprotein / Con A / receptor / photoaffinity label / blepharismone / 繊毛中 / ガモン / ブレファルモン / 糖タンパク質 / アスパラギン結合型糖鎖 / 有性生殖 / 繊毛虫の接合 / コンカナバリンA / カルシウムイオン / アスパラギンN-結合型 |
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| Research Abstract |
1. Studies on the structure of blepharmone (gamone 1)
Conjugation in Blepharisma japonicum is induced by interaction between complementary mating-types I and II, which excrete blepharmone (gamone 1) and blepharismone (gamone 2), respectively. Gamone 1 transforms type II cells such that they can unite, and gamone 2 similarly transforms type I cells. Gamone 1 was isolated and characterized as a glycoprotein of 20-30 kDa containing 175 amino acids and 6 sugars. To establish the structure of gamone 1, it was isolated by a new method based on concentration by Con A affinity column chromatography and purification by native PAGE. The partial amino acid sequence of four peptides obtained from gamone 1 was determined. By using mRNA from mating type, gamone 1 cDNA was isolated by rapid amplification. The cDNA sequence contains an ORF of 305 amino acids and codes a novel protein. We also estimated that the sugar sequence for the glycoprotein consisted from four N-acetylglucosamines and three mannoses.
2. Studies on the receptor of blepharismone (gamone 2)
To elucidate the receptor of blepharismone, a photoaffinity label probe of blepharismone is designed. Although the synthesis of it has not been attained so far, we synthesized a model compound N-benzoyl-blepharismone based on the strategy developed before. The model compound did not have mating inducing activity as expected, it was found to haye the significant activity of mating inhibition.
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