1998 Fiscal Year Final Research Report Summary
Development of protein crosslinking technology using the newly found family transglutaminase substrates termed cementoin
| Project/Area Number |
08558096
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Biomedical engineering/Biological material science
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| Research Institution | TOKYO INSTITUTE OF TECHNOLOGY |
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Principal Investigator |
SUZUKI Yoko Research Assistant Professor Tokyo Institute of Technology Faculty of Bioscience and Biotechnology, 生命理工学部, 教務職員 (30179270)
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| Co-Investigator(Kenkyū-buntansha) |
TACHIBANA Shinro Director, Eisai Co., Ltd., Department of Exploratory Drug Research II (Visiting, 研究開発本部, 部長(シンガポール大
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| Project Period (FY) |
1996 – 1998
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| Keywords | Crosslinking / Transglutaminase / Trappin / Cementoin / Green fluorescent protein / Gene conversion / Evolution / Glue |
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| Research Abstract |
To develop an efficient method for protein crosslinking that is necessary for the design of highly organized, multifunctional protein complexes, we carried out the following basic and biotechnological studies on the newly found transglutaminase substrates termed trappins. Trappins are a family of proteins that have anchoring sequences at their amino termini through which they become covalently trapped at their sites of action ; the anchoring sequence is called cementoin moiety. 1) Using recombinant cementoin moieties, we showed that their sequences rich in Gin, Lys, and Pro have a random coil structure and serve as a good substrate for transglutaminase, a characteristics very useful for developing the biotechnology of protein crosslinking or protein gluing. 2) SPAI, a circulating plasma trappin, was demonstrated to be synthesized and secreted by the enteroendocrine cells located near the crypt base in small intestine. 3) Fluorescent cementoin was prepared by fusing it with green fluorescent protein. The useful properties of the component proteins are maintained in the fusion protein, namely we succeeded to prepare a fluorescent cementoin moiety that will be of special interest and importance in protein engineering. 4) Evolution of cementoin family genes was clarified. Analysis of gene structure revealed that genes for trappins have evolved by the shuffling of domains between the REST and SLPI genes and that the intron sequences have been homogenized by gene conversion.
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