| Project/Area Number |
10557212
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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 | 展開研究 |
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| Research Field |
Physical pharmacy
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| Research Institution | The University of Tokyo |
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Principal Investigator |
IMAI Kazuhiro The University of Tokyo, Graduate School of Pharmaceutical Sciences, Professor, 大学院・薬学系研究科, 教授 (50012620)
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| Co-Investigator(Kenkyū-buntansha) |
FUKUSHIMA Takeshi The University of Tokyo, Graduate School of Pharmaceutical Sciences, Research associate, 大学院・薬学系研究科, 助手 (00272485)
SANTA Tomofumi The University of Tokyo, Graduate school of Pharmaceutical Sciences, Research associate, 大学院・薬学系研究科, 助手 (30187306)
HOMMA Hiroshi The University of Tokyo, Graduate School of Pharmaceutical Sciences, Associate Professor, 大学院・薬学系研究科, 助教授 (50190278)
FUJIWAKE Hideshi Shimazu Co. Ltd., Researcher, 研究員
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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 |
¥11,700,000 (Direct Cost: ¥11,700,000)
Fiscal Year 1999: ¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1998: ¥9,300,000 (Direct Cost: ¥9,300,000)
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| Keywords | sequensor / Edman degradation / fluorescent Edman reagent / fluorimetric HPLC / optical resolution / benzofurazan / ペプチドシークエンサー / PITC |
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| Research Abstract |
We have developed a sensitive method for the simultaneous determination of an amino acid sequence and D/L-configuration of peptides with fluorogenic Edman reagents and BFィイD23ィエD2 as cyclization/cleavage reagent.
First, New fluorogenic Edman reagents, PSBD-NCS and MSBD-NCS, were synthesized and the amino acids thiazolinone derivatives of these reagents were sensitively detected.
Secondly, we tried to detect more stable amino acids derivatives of these reagents. The amino acids thiazolininone derivatives of DBD-NCS were converted to carbamoyl derivatives via thiocarbamoyl compounds and detected fluorimetrically.
Thirdly, we investigated the conversion reaction of thiazolinone to thiohydantoin derivatives and found out that hydrochloric acid in methanol solution efficiently suppressed the racemization in the conversion reaction.
Finally, fluorogenic Edman reagent, MTBD-NCS, was synthesized. The thiocamamoyl and carbamoyl derivatives were not fluorescent and thiohydantoin derivatives of MTBD-NCS were fluorescent, giving the superiority with regard to avoidance from the interferring peaks of the degradation products.
The combined use of MTBD-NCS and BFィイD23ィエD2 gave us the sensitive determination method of amino acid sequence and D/L-configuration of peptides. This method developed in this study seems to be easily applicable to the conventional automatic sequensor.
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