Studies about protein structural alteration switching using in vivo mimic system.

使用体内模拟系统进行蛋白质结构改变转换的研究。

• 批准号: 14350433
• 负责人: TAKAGI Masahiro
• 金额: $ 9.34万
• 依托单位: JAPAN ADVANCED INSTITUTE OF SCIENCE AND TECHNOLOGY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14350433/
• 关键词: Amyloid; Fibril; Aggregation; Amine; Protein; アルギニン; ポリアミン; 熱成熟; 構造転移; 超好熱菌

Protein aggregation is facilitated by a partial unfolding during thermal or oxidative stress and byalterations in primary structure caused by mutation. Protein aggregation is a major problem in biomedical and biotechnological fields. In addition, with a growing expansion of genomic data, protein purification is one of the keys to perform post-genomic researches. In our living cells, protein aggregates are highly associated with fatal diseases, such as Alzheimer's, Creutzfeldt-Jakob, Huntington's and Parkinson's diseases. In this research, we focus on amine compounds as additive for preventing heat-induced aggregation of lysozyme, as a model protein. Furthermore, we investigated the fibrillogenesis of amyloid β and β_2-microglobulin in the presence of the additives.We investigated whether small amine compounds prevent heat-induced aggregation and inactivation of lysozyme and fibrilization of β_2-microglobulin. The following conclusions are proposed.(i)o-Diaminocyclohexane and p-phenylenediamine prevented heat-induced aggregation andinactivation of lysozyme. The inactivation rates of lysozyme in the presence of the additives were 3-fold slower than that in the absence of additives. (ii)p-Diaminocyclohexane and spermine prevented fibrilization of β_2-microglobulin. The rate of the fibrils in the presence of the additives is 4-fold slower than that in the absence of additives. These data provided new candidates for small additives to prevent protein aggregation.

热或氧化应激期间的部分解折叠以及突变引起的一级结构的改变促进了蛋白质聚集。蛋白质聚集是生物医学和生物技术领域的一个主要问题。此外，随着基因组数据的不断扩展，蛋白质纯化是其中之一。进行后基因组研究的关键在我们的活细胞中，蛋白质聚集体与致命疾病密切相关，例如阿尔茨海默病、克雅氏病、亨廷顿病和在这项研究中，我们重点关注胺类化合物作为模型蛋白来防止热诱导的溶菌酶聚集。此外，我们还研究了添加剂存在下β淀粉样蛋白和β_2-微球蛋白的纤维形成。小胺化合物能否阻止热诱导的溶菌酶聚集和失活以及β_2-微球蛋白的纤维化以下结论。 (i)邻二氨基环己烷和对苯二胺可防止热诱导的溶菌酶聚集和失活。在添加剂存在下，溶菌酶的失活速率比不存在添加剂时慢3倍。 -二氨基环己烷和精胺防止了β_2-微球蛋白的纤维化在添加剂存在下的纤维化率。这些数据为防止蛋白质聚集的小添加剂提供了新的候选者。

项目成果

Dissolution of protein aggregation by small amine compounds

小胺化合物溶解蛋白质聚集

• 发表时间: 2003
• 期刊: Science and Technology of Advanced Materials (STAM) 4
• 作者: Kentaro Shiraki;Motonori Kudou;Yoshinobu Aso;Masahiro Takagi.
• 通讯作者: Masahiro Takagi.

Screening for stable mutants with amino-acid pairs substituted for the disulfide bond between residues 14 and 38 of bovine pancreatic trypsin inhibitor (BPTI).

筛选用氨基酸对取代牛胰蛋白酶抑制剂 (BPTI) 残基 14 和 38 之间的二硫键的稳定突变体。

• 发表时间: 2002
• 期刊: J.Biol.Chem. 277
• 作者: Yoshihisa Hagihara;Kentaro Shiraki;Tsutomu Nakamura;Koichi Uegaki;Masahiro Takagi;Tadayuki Imanaka;Noboru Yumoto
• 通讯作者: Noboru Yumoto

Mutational effects on 06-methylguanine-DNA methyltransferase from hyperthermophile contribution of ion-pair network to protein thermostability.

离子对网络的超嗜热菌对蛋白质热稳定性的贡献对 06-甲基鸟嘌呤-DNA 甲基转移酶的突变影响。

• 发表时间: 2004
• 期刊: J.Biochemistry(Tokyo) 135
• 作者: S.Nishikori et al.

Mutational effects on O6-methylguanine-DNA methyltransferase from hyperthermophile contribution of ion-pair network to protein thermostability.

离子对网络的超嗜热菌对蛋白质热稳定性的贡献对 O6-甲基鸟嘌呤-DNA 甲基转移酶的突变影响。

• 发表时间: 2004
• 期刊: J.Biochemistry (Tokyo) 135
• 作者: An;C.-I.ほか;S.Nishikori et al.
• 通讯作者: S.Nishikori et al.

Arginine ethylester prevents thermal inactivation and aggregation of lysozyme.

• DOI: 10.1111/j.1432-1033.2004.04257.x
• 发表时间: 2004-08
• 期刊: European journal of biochemistry
• 作者: K. Shiraki;M. Kudou;S. Nishikori;Harue Kitagawa;T. Imanaka;M. Takagi