| Project/Area Number |
09460055
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Bioproduction chemistry/Bioorganic chemistry
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| Research Institution | The University of Tokyo |
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Principal Investigator |
NAGASAWA Hiromichi The University of Tokyo, Graduate School of Agricultural and life Sciences, Department of Applied Biological Chemistry, Professor, 大学院・農学生命科学研究科, 教授 (60134508)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥7,300,000 (Direct Cost: ¥7,300,000)
Fiscal Year 1998: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1997: ¥5,300,000 (Direct Cost: ¥5,300,000)
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| Keywords | Crustaceans / Androgenic gland hormone / sex differentiation / amino acid sequence / sex reversal / Armadillidium vulgare / isopod / androgenic gland / 等脚類 |
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| Research Abstract |
Sex differentiation in crustaceans is known to be hormonally controlled. The androgenic glands develop only in males and produces a peptidic hormone called androgenic gland hormone (AGH), which promotes male character. AGH induces sex-reversal when injected into a young female. We have been purifying AGH from the androgenic glands of the terrestrial isopod, Armadillidium vulgare. Inactivation experiments revealed that AGH is heat-stable and peptidic, and that it contains disulfide bridge(s). Its molecular weight was estimated to be 11,000-13,000 by gel-filtration. AGH was extracted with buffer solution, and the extract was applied successively to ion-exchange HPLC and two steps of reverse-phase HPLC.Finally, we obtained highly purified AGH with sex-reversal activity at a dose of 38 pg. N-terminal amino acid sequence analysis afforded two kinds of PTH-amino acid at most of the cycles of Edman degradation. Mass spectral and SDS-PAGE analyses of the purified material, however, suggested that it was still impure. Then, AGH was purified again by the same way from a new batch of the androgenic glands and subjected to reductive carboxymethylation. Only one peptide was recovered. Sequence analysis of this peptide revealed a single sequence which was estimated to be one of the two sequences described above. These results suggested that AGH consisted of two peptide chains connected by disulfide bridges. This was supported by the sequence analysis of the purified sample from another batch of extraction source, which showed very similar results. Cloning of a cDNA encoding AGH is now in progress.
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