8G13

CryoEM structure of cytosolic GAPDH under 8h Oxidative Stress

8G13 の概要

• エントリーDOI: 10.2210/pdb8g13/pdb
• EMDBエントリー: 29660
• 分子名称: Glyceraldehyde-3-phosphate dehydrogenase (1 entity in total)
• 機能のキーワード: gapdh, energy, cytosolic protein, transferase
• 由来する生物種: Homo sapiens (Human)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 143871.88

構造登録者

Choi, W.Y.,Wu, H.,Cheng, Y.F. (登録日: 2023-02-01, 公開日: 2023-12-27)

主引用文献

Choi, W.,Wu, H.,Yserentant, K.,Huang, B.,Cheng, Y.
Efficient tagging of endogenous proteins in human cell lines for structural studies by single-particle cryo-EM.
Proc.Natl.Acad.Sci.USA, 120:e2302471120-e2302471120, 2023

PubMed Abstract: CRISPR/Cas9-based genome engineering has revolutionized our ability to manipulate biological systems, particularly in higher organisms. Here, we designed a set of homology-directed repair donor templates that enable efficient tagging of endogenous proteins with affinity tags by transient transfection and selection of genome-edited cells in various human cell lines. Combined with technological advancements in single-particle cryogenic electron microscopy, this strategy allows efficient structural studies of endogenous proteins captured in their native cellular environment and during different cellular processes. We demonstrated this strategy by tagging six different human proteins in both HEK293T and Jurkat cells. Moreover, analysis of endogenous glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in HEK293T cells allowed us to follow its behavior spatially and temporally in response to prolonged oxidative stress, correlating the increased number of oxidation-induced inactive catalytic sites in GAPDH with its translocation from cytosol to nucleus.

PubMed: 37487103
DOI: 10.1073/pnas.2302471120

実験手法

ELECTRON MICROSCOPY (2.3 Å)