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.

新发现的转谷氨酰胺酶底物，称为水泥素和水泥素样部分，是使用重组 DNA 技术在大肠杆菌中产生的，作为与麦芽糖结合蛋白的融合蛋白，通过淀粉糖树脂上的亲和层析进行纯化，并进行表征。该产品很容易与催化量的转谷氨酰胺酶交联，表明它们确实具有用作蛋白质交联剂或蛋白质共价胶的合适特性。还制备了特异性抗血清并用于蛋白质印迹分析、免疫组织化学染色和放射免疫分析；人们发现气管和肠的上皮细胞中富含交联蛋白。银屑病患者中水泥蛋白/elafin 的表达显着升高。这些结果表明，水泥素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且有用的分子。

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。”