構造学的解析に基づくＬＧＩ１－ＡＤＡＭ２２複合体形成の生理機能の解明

基于结构分析阐明LGI1-ADAM22复合物形成的生理功能

• 批准号: 17J07958
• 负责人: 宮崎 裕理
• 金额: $ 1.09万
• 依托单位: National Institute for Physiological Sciences (2018)The Graduate University for Advanced Studies (2017)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for JSPS Fellows
• 财政年份: 2017
• 资助国家: 日本
• 起止时间: 2017-04-26 至 2019-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17J07958/
• 关键词: LGIファミリー; 立体構造予測; タンパク質間相互作用; てんかん; LGI1; ADAM22; X線結晶構造解析; シナプス間架橋; てんかん変異モデルマウス

平成30年度の研究計画に従い、これまでに明らかにしたLGI1-ADAM22の詳細な結合様式を基に、種々の脳神経疾患の発症との関連性が報告されているLGI1のファミリータンパク質（LGI2-4）とADAM22の結合特異性の評価を行った。まず、マウス脳組織を利用した免疫沈降実験によって、ADAM22と生理的に結合しうるLGIファミリータンパク質を検証した。その結果、LGI1とLGI4は効率的にADAM22と結合したが、LGI2とLGI3はほとんど結合しないことを見出した。また、in vitroの結合実験によって、ADAM22に対するLGIファミリータンパク質の結合性を比較したところ、同様の結果が得られた。この結合性の違いを構造学的に検証するために、これまでに明らかにしたLGI1の結晶構造を鋳型としたLGIファミリータンパク質の3次元構造予測（ホモロジーモデリング）を行った。モデル構造を精査した結果、LGIファミリーは互いによく似た立体構造を有する一方で、ADAM22との結合面に相当するタンパク質の分子表面には、数アミノ酸残基の違いがあると予想された。そこで、このアミノ酸残基の違いによるADAM22との結合への影響を、ファミリータンパク質の変異体を使った結合実験により検証した。その結果、LGIファミリーの分子表面の僅かなアミノ酸残基の違いが、ADAM22との結合に決定的な違いを与えることを見出した。これらの結果は、LGIファミリータンパク質の生理的な機能の違いが、ADAM22サブファミリータンパク質との結合の違いに依存している可能性を示している。したがって、LGI2とLGI3は他の受容体分子との結合を介して機能する可能性が高いと考えられた。以上の結果は、LGIファミリータンパク質の機能の違いおよび、関連する脳神経疾患の発症機序を理解する上で、重要な基礎資料となると考えられた。

In Pingcheng's 30-year research program, the research plan shows that the combination of the LGI1-ADAM22 test and the serial report on the relationship between the LGI1 and the disease (LGI2-4) is a combination of the ADAM22 and the special practice. The combination of immunoprecipitation and ADAM22 physiology was used in the two groups. The combination of immune sedimentation and LGI physiology. The result of the experiment, the ADAM22 combination of the LGI1 LGI4 error rate, and the LGI2 LGI3 immune response were combined with the LGI2 LGI3 immune response. The results of the combination of in vitro and in vitro were compared, and the results of the same results were compared. The results showed that the combination of the two methods was better than that of the others, and the results of the combination of the two methods were compared with each other, and the results of the same test were compared with each other. The results showed that the results of the test were better than that of the other two, and the results of the same test were compared. The results showed that there were significant differences between the two groups. In this paper, the results of computer science are introduced, and the results show that the structure of the LGI1 system is based on the data of the computer science system. The results show that the three-dimensional structure of the LGI system is generated in the system. The results showed that there was a good relationship between LGI and ADAM22, and that the surface of the molecule was much higher than that on the surface of the molecule. Make sure that the acid residue and ADAM22 are combined with each other so that it can be combined with each other. The results showed that the molecular surface of LGI was stained with acid residues, and ADAM22 was combined with the determined results. The results of the experiment, the results of the LGI test, the ability of the physiological machine to perform the test, and the possibility of the combination of the results of the test and the combination of the results of the test and the combination of the results of the experiment, the physiological response of the physiology, and the probability of the combination of the results of the experiment, the possibility of the combination of the results and the results of the ADAM22 test, the possibility of the combination of the results of the test and the probability of the combination of the results of the test, the physiological response of the physiology, and the possibility of the combination of the results of the test, the possibility of the combination of the results and the results of the test, the possibility of the combination of the results of the test, the results of the test, the possibility of the combination of the results of the test, the possibility of the combination of The combination of LGI2, LGI2 and other capacitive molecules in combination with the introduction of high probability of mechanical testing is very important. According to the above results, LGI is able to detect the symptoms of disease, understand the sequence of symptoms of diseases and important basic information.

项目成果

Binding mode and physiological role of epilepsy-related ligand/receptor LGI1-ADAM22 complex based on structural analysis

基于结构分析的癫痫相关配体/受体LGI1-ADAM22复合物的结合模式和生理作用

• 发表时间: 2018
• 作者: Yamagata Atsushi;Miyazaki Yuri;Yokoi Norihiko;Shigematsu Hideki;Sato Yusuke;Goto-Ito Sakurako;Maeda Asami;Goto Teppei;Sanbo Makoto;Hirabayashi Masumi;Shirouzu Mikako;Fukata Yuko;Fukata Masaki;Fukai Shuya;Yuri MIYAZAKI
• 通讯作者: Yuri MIYAZAKI