Structural fluctuations of protein complexes measured by combination of protein engineering and solvent quenching meshod

蛋白质工程和溶剂淬灭技术相结合测量蛋白质复合物的结构波动

• 批准号: 23770194
• 负责人: MAKABE koki
• 金额: $ 2.91万
• 依托单位: Yamagata University (2012-2013)National Institutes of Natural Sciences Okazaki Research Facilities (2011)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2013
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23770194/
• 关键词: タンパク質工学; シャペロン; 水素交換; GroEL; 蛋白質; 構造揺らぎ; タンパク質; 水素重水素交換

In this project, we have been studying the structural fluctuations of E. coli chaperonin GroEL/ES system by using DMSO-quenched H/D exchange methods. For GroES H/D exchange experiments, we have successfully determined the exchange rates for 33 amide protons out of 94 amide protons in GroES. For the remaining 61 amide protons, we have determined the lower limit of the exchange rates. We found that only 10 amide protons have a protection factor larger than 10 to the power 6th. This result indicates that the free GroES is highly flexible in significant portions. From crystal structure analysis of N-terminal mutants of alpha-lactalbumin, a substrate protein of GroEL/ES system, we found the hydrogen bonding of the N-terminal residue is important for its stability.

在这个项目中，我们一直在研究E的结构波动。coli伴侣蛋白GroEL/ES系统。对于GroES H/D交换实验，我们已经成功地确定了GroES中94个酰胺质子中的33个酰胺质子的交换率。对于剩余的61个酰胺质子，我们已经确定了交换率的下限。我们发现只有10个酰胺质子的保护因子大于10的6次幂。该结果表明，自由GroES在显著部分中是高度柔性的。通过对GroEL/ES系统的底物蛋白α-乳白蛋白N端突变体的晶体结构分析，发现N端残基的氢键对其稳定性有重要影响。

项目成果

蛋白質-脂肪酸複合体及び腫瘍細胞の細胞死誘導薬

蛋白质-脂肪酸复合物和肿瘤细胞死亡诱导剂

• 发表时间: 2011

Sequential four-state folding/unfolding of goat α-lactalbumin and its N-terminal variants.

山羊 α-乳清蛋白及其 N 末端变体的连续四态折叠/展开。

• DOI: 10.1002/prot.24109
• 发表时间: 2012
• 期刊: Proteins.
• 作者: Tomoyori K;Nakamura T;Makabe K;Maki K;Saeki K;Kuwajima K.
• 通讯作者: Kuwajima K.

STRUCTURAL INSIGHTS INTO THE STABILITY PERTURBATIONS BY N-TERMINAL VARIATIONS IN HUMAN AND GOAT ALPHA-LACTALBUMIN

人类和山羊 α-乳白蛋白 N 端变异引起的稳定性扰动的结构见解

• 发表时间: 2012
• 作者: Koki Makabe;Takashi Nakamura and Kunihiro Kuwajima
• 通讯作者: Takashi Nakamura and Kunihiro Kuwajima

Sequential Four-state Folding/unfolding of Goatα-Lactalbumin and Its N-Terminal Variants Proteins

山羊α-乳清蛋白及其 N 端变体蛋白的连续四态折叠/解折叠

• 发表时间: 2012
• 作者: Tomoyori K;Nakamura T;Makabe K;Maki K;Saeki K;Kuwajima K
• 通讯作者: Kuwajima K

Folding and redesign of a beta-sheet rich model protein

富含β折叠的模型蛋白的折叠和重新设计

• 发表时间: 2013
• 作者: Koki Makabe