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.

蛋白质聚集是通过在热应力或氧化应激期间的部分展开来制备的，而由突变引起的一级结构中的伴侣。蛋白质聚集是生物医学和生物技术领域的主要问题。此外，随着基因组数据的扩大，蛋白质纯化是进行基因组研究后研究的关键之一。在我们的活细胞中，蛋白质骨料与致命疾病高度相关，例如阿尔茨海默氏症，克鲁特兹菲尔德·贾科布，亨廷顿和帕金森氏病。在这项研究中，我们专注于胺化合物作为预防热诱导的溶菌酶作为模型蛋白的添加剂。此外，我们研究了在添加剂的存在下淀粉样β和β_2-微球蛋白的原纤维生成。我们研究了小胺化合物是否可以阻止热诱导的聚集和溶菌酶失活和β_2-微球蛋白的纤维化和纤维化。提出了以下结论。（i）o-二氨基细胞环己烷和p-苯二胺可阻止热诱导的聚集和溶菌酶激活。在添加剂存在下，溶菌酶的灭活率比没有添加剂的情况慢3倍。 （ii）p-二氨基辛己烷和精子可防止β_2-微球蛋白的纤维化。在添加剂存在下，原纤维的速率比没有添加剂的情况慢4倍。这些数据为小型添加剂提供了新的候选，以防止蛋白质聚集。

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