2007 Fiscal Year Final Research Report Summary
Application of the intra-molecular chaperon function of propeptide of peptide hormone for de novo design of bioactive peptide
| Project/Area Number |
16510160
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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 |
Living organism molecular science
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| Research Institution | Kinki University |
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Principal Investigator |
HIDAKA Yuji Kinki University, Life Science, Associate Professor (70212165)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAGUCHI Hiroshi Kwanseigakuin University, Chemistry, Associate Professor (10252719)
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| Project Period (FY) |
2004 – 2007
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| Keywords | intra-molecular chaperone / precursor / peptide hormone / propeptide / folding / uroguanylin / processing |
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| Research Abstract |
A pro-peptide of uroguanylin functions as an intra-molecular chaperone for the correct folding of the mature peptide, uroguanylin. In order to further study the intra-molecular chaperone function, a series of de novo designed peptides, of which tertiary structures were kinetically trapped by the pro-peptide of uroguanylin, was prepared and its biological activity and folding.
In order to investigate the structural basis of the chaperone function, pro-uroguanylin was prepared by E. coli expression system. Crystal screening was performed using the crystal screen kit of Hampton research. Crystals of pro-uroguanylin was applied for the collection of reflections in Spring 8. The phase determination is now in progress.
The intra-molecular function of propeptide was applied for the de novo design of bioactive peptide. For this purpose, a series of disulfide hybrid peptides of uroguanylin and heat-stable enterotoxin (ST) was designed and chemically synthesized. The fusion proteins of propeptide of uroguanylin and the hybrid peptides were also prepared by E. coli expression system. In order to estimate whether the propeptide is able to kinetically trap the hybrid peptides, folding reactions of hybrid peptides and fusion proteins were examined. We obtained a novel bioactive peptide, which was kinetically trapped by the propeptide, using this screening system. In order to obtain the structural information, CD measurements were performed. The hybrid peptide showed a CD spectrum similar to that of ST and uroguanylin.
The results showed that the only bioactive peptide can be trapped by the propeptide, suggesting that the tertiary structure of the interacting space in the propeptide provides the template for the bioactive structure.
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