Developemt of protein crosslinking technology using the newly found transglutaminase substrate termed cementoin.

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The newly found transglutaminase substrates, termed cementoin and cementoin-like moieties, were produced in E.coli using recombinant DNA technology as fusion proteins with maltose-binding protein, purified by affinity chromatography on amyrose resin, and characterized. The products were readily cross-linked with a catalytic amount of transglutaminase, indicating that they have indeed appropriate properties to be used as a protein cross-linker or protein covalent glue. Specific antisera were also produced and used for Western blot analysis, immunohistochemical staining, and radioimmunoassay ; the cross-linker proteins were found to be rich in the epitherium of trachea and intestine. Expression of cementoin/elafin was greatly elevated in a psoriasis patient. These results indicate that cementoin and its homologues are important and useful molecules not only in the field of protein engineering but also in understanding basic physiology.