| Project/Area Number |
09672152
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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 |
Chemical pharmacy
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| Research Institution | Health Sciences University of Hokkaido |
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Principal Investigator |
TANIZAWA Kazutaka Health Sciences University of Hokkaido, Faculty of Pharm.Sci., professor, 薬学部, 教授 (90001049)
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| Co-Investigator(Kenkyū-buntansha) |
TOYOTA Eiko Health Sciences University of Hokkaido, Faculty of Pharm.Sci., lecturer, 薬学部, 講師 (00103200)
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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 |
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1998: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1997: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | trypsin / affinity labeling / trypsin-like enzyme / chemical modification / metal chelate / peptide bond cleavage / トリプシン / トリプシン類縁酵素 / ペプチド切断試薬 |
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| Research Abstract |
In our previous work, it was found that benzamidine derivatives behave as strong binding affinity towards trypsin and trypsin-like enzymes. As an extension of the wurk, we designed compounds which contained metal chelate moiety as well as the amidine group with a view to obtaining selective modification reagents for trypsin and trypsin-like enzymes. The reagents were expected to mediate cleavage of the enzyme backbone at the region where the metal attained close contact when they were bound in the binding site of the enzyme. the three-dimensional structure of the enzyme-reagent complex. eaction peptide bond cleavage reagents for trypsin.
In 1997 we synthesized 38 chelates as candidate for trypsin-specific cleavage reagent, and their binding affinity toward trypsin and trypsin-like enzymes was determinecL They were analyzed to exhibit strong binding affinity toNard these enzymes with Ki values range of 10^5 - 1O^6 Mi. During the period of the fiscal year of 1998 the medification reaction mediated by these chelates were investigated. Cleavage was observed when the enzyme was incubated with the reagent in the presence of hydrogenperoxide and ascorbate. A peptide fragment due to bond cleavage was analyzed by SDS- PAGE.It was suggested that the cleavage occurred at the region within Gly195 -A1a2O4. It was concluded that the region is close to the active site of the enzyme in its three-dimensional structure.
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