論文

基本情報

氏名 中村 元直
氏名(カナ) ナカムラ モトナオ
氏名(英語) Nakamura Motonao
所属 生命科学部 生物科学科
職名 教授
researchmap研究者コード 6000014705
researchmap機関 岡山理科大学

題名

The leukotriene B4 receptor BLT1 is stabilized by transmembrane helical capping mutations.

単著・共著の別

著者

Hori,T., Nakamura,M., Yokomizo,T., Shimizu,T., and Miyano,M.

概要

In this study, we introduced structure-based rational mutations in the guinea pig leukotriene B4 receptor
(gpBLT1) in order to enhance the stabilization of the protein. Elements thought to be unfavorable for the
stability of gpBLT1 were extracted based on the stabilization elements established in soluble proteins,
determined crystal structures of G-protein-coupled receptors (GPCRs), and multiple sequence alignment. The two unfavorable residues His832.67 and Lys883.21, located at helix capping sites, were replaced with Gly (His83Gly2.67 and Lys88Gly3.21). The modi ed protein containing His83Gly2.67/Lys88Gly3.21 was highly expressed, solubilized, and puri ed and exhibited improved thermal stability by 4°C in comparison with that of the original gpBLT1 construct. Owing to the double mutation, the expression level increased by 6-fold (Bmax ¼311 pmol/mg) in the membrane fraction of Pichia pastoris. The ligand binding afnity was similar to that of the original gpBLT1 without the mutations. Similar unfavorable residues have been observed at helix capping sites in many other GPCRs; therefore, the replacement of such residues with more favorable residues will improve stabilization of the GPCR structure for the crystallization.

発表雑誌等の名称

Biochemistry and Biophysics Reports

出版者

開始ページ

終了ページ

発行又は発表の年月

2015/11

査読の有無

有り

招待の有無

無し

記述言語

英語

掲載種別

研究論文(学術雑誌)

ISSN

ID:DOI

ID:NAID(CiNiiのID)

ID:PMID

URL

JGlobalID

arXiv ID

ORCIDのPut Code

DBLP ID