Analysis of protein folding memory with modification of intramolecular chaperons and its application

分子内伴侣修饰的蛋白质折叠记忆分析及其应用

• 批准号: 24656501
• 负责人: UEDA Mitsuyoshi
• 金额: $ 2.58万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2012
• 资助国家: 日本
• 起止时间: 2012 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-24656501/
• 关键词: プロテインフォールディングメモリー; プロペプチド; 分子内シャペロン; カルボキシペプチダーゼ Y; 分子ディスプレイ; 活性化エネルギー; アンフィンセン理論; 触媒効率; カルボキシペプチダーゼY

CPYs (carboxypeptidase Y) with mutated propeptides were successfully displayed on yeast cell surface, and the mature enzymes were purified by the selective cleavage of mutated propeptides. Measurement of the activity and kinetics of the displayed CPYs indicated that the propeptide mutation altered the catalytic efficiency of mCPY. Although the mature region of the wild -type and mutant CPYs had identical amino acid sequences, the mCPYs from the mutant proCPYs had higher catalytic efficiency than the wild-type. These results indicate that proteins with identical amino acid sequence can fold into isomeric proteins with micro-conformational changes through mutated intramolecular chaperones.

将具有突变前肽的羧肽酶Y （carboxypeptidase Y）成功地展示在酵母细胞表面，并通过对突变前肽的选择性切割纯化成熟酶。对所展示的cppy的活性和动力学测量表明，前肽突变改变了mCPY的催化效率。虽然野生型和突变型CPYs的成熟区氨基酸序列相同，但突变型proCPYs的mCPYs具有比野生型更高的催化效率。这些结果表明，具有相同氨基酸序列的蛋白质可以通过分子内伴侣的突变折叠成具有微构象变化的异构体蛋白质。

项目成果

タンパク質フォールデイングメモリーによる酵素の改変

通过蛋白质折叠记忆修饰酶

• 发表时间: 2012
• 作者: M. Nagayama;H. Maeda;K. Kuroda;M. Ueda;里村淳ら
• 通讯作者: 里村淳ら

Identification of Interaction Site of Propeptide toward Mature Carboxypeptidase Y (mCPY) Based on the Similarity between Propeptide and CPY Inhibitor (IC)

• DOI: 10.1271/bbb.110668
• 发表时间: 2012-01
• 期刊: Bioscience, Biotechnology, and Biochemistry
• 作者: M. Nagayama;K. Kuroda;M. Ueda