2014 Fiscal Year Final Research Report
Mechanism of FeS cluster-binding crosslink enzyme and production of novel cyclic peptide
| Project/Area Number |
24658288
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Applied molecular and cellular biology
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| Research Institution | Osaka University |
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Principal Investigator |
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| Project Period (FY) |
2012-04-01 – 2015-03-31
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| Keywords | 鉄硫黄クラスター / チオエーテル架橋 / 環状ペプチド |
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| Outline of Final Research Achievements |
An operon coding quinohemoprotein amine dehydrogenase (QHNDH) contains an ORF2 protein that generates three intra-peptidyl thioether crosslinks between Cys and Asp/Glu residues in the gamma subunit of QHNDH. As a result, 5-8 residues of loop regions were formed in the multi-loop structure of the gamma subunit. To elucidate the reaction mechanism of the protein and to construct peptide library cyclized by thioether bond formation, the ORF2 protein was purified under anaerobic conditions. Reconstitution of FeS cluster provided the active form catalyzing thioether bond formation in vitro. On the basis of the homology model of the ORF2 complexed with the gamma subunit, the details of the reaction mechanism were revealed in this study. In the short type gamma subunit containing a single crosslink site, it is successful that various sequences as well as randomized sites are introduced to the loop region.
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| Free Research Field |
生化学
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