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

使用新发现的转谷氨酰胺酶底物（称为水泥素）开发蛋白质交联技术。

• 批准号: 05558109
• 负责人: HIROSE Shigehisa
• 金额: $ 4.8万
• 依托单位: TOKYO INSTITUTE OF TECHNOLOGY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05558109/
• 关键词: Crosslinking; Transglutaminase; Elafin; Glue; Cementoin; バイオコンジュゲート; ハイブリッド; タンパク質の接着; 内在性接着因子

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.

利用重组DNA技术作为麦芽糖结合蛋白的融合蛋白，在大肠杆菌中生产了新发现的谷氨酰胺转氨酶底物，命名为胶原蛋白和类胶原蛋白，经淀粉糖树脂亲和层析纯化，并对其进行了鉴定。产物很容易与催化量的转谷氨酰胺酶交联，表明它们确实具有作为蛋白质交联剂或蛋白质共价胶的适当性质。还制备了特异性抗血清，用于Western印迹分析、免疫组织化学染色和放射免疫分析，发现交联物蛋白在气管和肠道上皮中含量丰富。1例银屑病患者外周血中牙骨质/弹力蛋白的表达显著升高。这些结果表明，牙胶蛋白及其同系物不仅在蛋白质工程领域，而且在理解基础生理学方面都是重要和有用的分子。

项目成果

Ichiro Tamechika et.al.: "Accelerated evolution in the cementoin and inhibitor domains of porcine elafin family members." 271, (in press).

Ichiro Tamechika 等人：“猪弹性蛋白家族成员的水泥素和抑制剂结构域的加速进化。”

Kyoko Nonomura, et.al.: "Up-regulation of elafin/SKULP gene expression in psoriatic epidermis." J.Invest.Dermatol.103. 88-91 (1994)

Kyoko Nonomura 等人：“银屑病表皮中 elafin/SKULP 基因表达的上调。”

Hiromi Hagiwara, et.al.: "Immunochemical characterization and localization of endothelin ETB receptor." Am.J.Physiol.264. F777-F783 (1994)

Hiromi Hagiwara 等人：“内皮素 ETB 受体的免疫化学特征和定位。”

Nara,K.: "Elastase inhibitor elafin is a new type of proteinase inhibitor which has a transglutaminese-mediated anchoring sequence termed “Cementoin"" J.Biochem.115. 441-448 (1994)

Nara, K.：“弹性蛋白酶抑制剂 elafin 是一种新型蛋白酶抑制剂，具有转谷氨酰胺酶介导的锚定序列，称为“Cementoin””J.Biochem.115 (1994)。

Kyoko Nonomura: "Up-regulation of elafin/SKALP gene expression in psoriatic epidermis.21GC03:J.Invest.Dermatol." 103. 88-91 (1994)

Kyoko Nonomura：“银屑病表皮中 elafin/SKALP 基因表达的上调。21GC03：J.Invest.Dermatol。”