Dynamics of protein structure alteration at single molecular level

单分子水平蛋白质结构改变的动力学

• 批准号: 17360395
• 负责人: TAKAGI Masahiro
• 金额: $ 10.36万
• 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: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17360395/
• 关键词: Protein; Structure; Amyloid; Membrane; Interaction; タンパク質; フォールディング; 構造変化; 拡張遺伝子工学; ウシトリプシンインヒビター; ミクログロブリン

(1) In order to perform analysis of protein structural alteration at single molecular level using FRET method We attempted fluorescence labeling of bavine pancreatic tripsin inhibitor (BFTI) using a new technique of four base codon method. Fluorescent proteins were obtained but not enough yield for studies at molecular level(2) The effects of negatively charged phosphatidylserine (PS) and neutral phosphatidylcholine (PC) on the structure of wild-type and mutant bovine pancreatic trypsin inhibitor (BPTI) at neutral pH were investigated. The presence of PC did not have any effect on the protein structure whilst PS induced a non-native structure in three mutant BPTI proteins. However, the negatively charged phospholipid did not have any effect on wild-type BPTI. The findings revealed that (i) elimination of some disulphide bonds, results in dramatic change in protein structure, and, (ii) that this biochemical interaction is surface-driven and that electrostatic interactions may play a ver … More y strong role in influencing the fore-stated changes in protein structure.(3) Patients on long term hemodyalysis frequently suffer from dialysis-related amyloidosis (DRA). DRA is a pathology associated with a persistent increase of monomeric β2- microglobulin (β2-m) caused by dialysis treatment of renal failure. The mechanism of formation of amyloid fibrils for this protein, as for all the other amyloidogenic proteins, is not clear, and there was great uncertainty about the molecular bases of amyloid deposition when the amyloidogenicity of β2-m was discovered. In this study, we investigated formation of variant β2-m in serum of a patient who receives treatment of hemodialysis. Variant β2-m is derived from restricted cleavage of β2-M. The possible factor involved in formation of variant β2-m was also estimated. The following conclusions are proposed. Formation of variant β2-m, namely restricted cleavage of β2-m, might be caused by proteolytic cleavage by a serum protease(s). However, it is not clear whether the variant β2-m can trigger formation of amyloid. Protein aggregation formation is enhanced in serum from a patient. Less

（1）为了使用FRET方法对单分子水平进行蛋白质结构改变，我们尝试使用四种基本密码子方法的新技术对Bavine Ancreatic Tripsin抑制剂（BFTI）进行荧光。对于分子水平的研究（2）负电荷磷脂酰丝氨酸（PS）和中性磷脂酰胆碱（PC）Wil d型和突变牛胰腺胰蛋白酶抑制剂（BPTI）对中性pH有任何影响三种Mutti蛋白中的非本地结构对野生型BPTI没有任何影响。驱动和静电在影响蛋白质结构的前期变化（3）患者的长期血流分析中的强大作用。透析治疗肾脏衰竭的机制是蛋白质的透析机制，因此在这项研究中发现了β2-M的淀粉样蛋白。血液透析是从形成的β2-m的。血清蛋白酶可触发蛋白质聚集的形成

项目成果

Endoplasmic reticulum stress responses as criteria to evaluate functional foods and cosmetics

内质网应激反应作为评价功能性食品和化妆品的标准

• 发表时间: 2007
• 作者: M. Takagi;K. Yamaguchi;T. Hamada (Invited)
• 通讯作者: T. Hamada (Invited)

光による脂質二分子膜小胞の形態転移

光诱导脂质双层囊泡的形状转变

• 发表时间: 2007
• 作者: 石井健一;長崎健;浜田勉;高木昌宏
• 通讯作者: 高木昌宏

生体モデル膜ドメインの出芽ダイナミクス

生物模型膜域的萌芽动力学

• 发表时间: 2007
• 作者: 浜田勉;高木昌宏;三浦陽子
• 通讯作者: 三浦陽子

Studies about stability of membrane raft domain

膜筏结构域稳定性研究

• 发表时间: 2007
• 作者: Yamaguchi;K.;et. al.
• 通讯作者: et. al.

Unfolding Mechanism of a Hyperthermophilic Protein 06-methylguanine-DNA Methyltransferase

超嗜热蛋白 06-甲基鸟嘌呤-DNA 甲基转移酶的解折叠机制

• 发表时间: 2005
• 期刊: Biophysical Chemistry 116
• 作者: Nishikori;S.;Shiraki;K.;Fujiwara;S.;Imanaka;T.;Takagi;M
• 通讯作者: M