5Z4B

GB1 structure determination in living eukaryotic cells by in-cell NMR spectroscopy

5Z4B の概要

• エントリーDOI: 10.2210/pdb5z4b/pdb
• NMR情報: BMRB: 36153
• 分子名称: Protein LG (1 entity in total)
• 機能のキーワード: protein, immune system
• 由来する生物種: Finegoldia magna (Peptostreptococcus magnus)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 6258.84

構造登録者

Tanaka, T.,Teppei, I.,Kamoshida, H.,Mishima, M.,Shirakawa, M.,Guentert, P.,Ito, Y. (登録日: 2018-01-10, 公開日: 2019-01-23, 最終更新日: 2024-05-15)

主引用文献

Tanaka, T.,Ikeya, T.,Kamoshida, H.,Suemoto, Y.,Mishima, M.,Shirakawa, M.,Guntert, P.,Ito, Y.
High-Resolution Protein 3D Structure Determination in Living Eukaryotic Cells.
Angew.Chem.Int.Ed.Engl., 58:7284-7288, 2019

PubMed Abstract: Proteins in living cells interact specifically or nonspecifically with an enormous number of biomolecules. To understand the behavior of proteins under intracellular crowding conditions, it is indispensable to observe their three-dimensional (3D) structures at the atomic level in a physiologically natural environment. We demonstrate the first de novo protein structure determinations in eukaryotes with the sf9 cell/baculovirus system using NMR data from living cells exclusively. The method was applied to five proteins, rat calmodulin, human HRas, human ubiquitin, T. thermophilus HB8 TTHA1718, and Streptococcus protein G B1 domain. In all cases, we could obtain structural information from well-resolved in-cell 3D nuclear Overhauser effect spectroscopy (NOESY) data, suggesting that our method can be a standard tool for protein structure determinations in living eukaryotic cells. For three proteins, we achieved well-converged 3D structures. Among these, the in-cell structure of protein G B1 domain was most accurately determined, demonstrating that a helix-loop region is tilted away from a β-sheet compared to the conformation in diluted solution.

PubMed: 30938016
DOI: 10.1002/anie.201900840

実験手法

SOLUTION NMR