7VDC

3.28 A structure of the rabbit muscle aldolase

7VDC の概要

• エントリーDOI: 10.2210/pdb7vdc/pdb
• EMDBエントリー: 31913
• 分子名称: Fructose-bisphosphate aldolase A (1 entity in total)
• 機能のキーワード: rabbit muscle aldolase, structural protein
• 由来する生物種: Oryctolagus cuniculus (Rabbit)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 157579.50

構造登録者

Fan, H.C.,Zhang, Y.,Sun, F. (登録日: 2021-09-06, 公開日: 2021-12-29, 最終更新日: 2025-06-25)

主引用文献

Fan, H.,Wang, B.,Zhang, Y.,Zhu, Y.,Song, B.,Xu, H.,Zhai, Y.,Qiao, M.,Sun, F.
A cryo-electron microscopy support film formed by 2D crystals of hydrophobin HFBI.
Nat Commun, 12:7257-7257, 2021

PubMed Abstract: Cryo-electron microscopy (cryo-EM) has become a powerful tool to resolve high-resolution structures of biomacromolecules in solution. However, air-water interface induced preferred orientations, dissociation or denaturation of biomacromolecules during cryo-vitrification remains a limiting factor for many specimens. To solve this bottleneck, we developed a cryo-EM support film using 2D crystals of hydrophobin HFBI. The hydrophilic side of the HFBI film adsorbs protein particles via electrostatic interactions and sequesters them from the air-water interface, allowing the formation of sufficiently thin ice for high-quality data collection. The particle orientation distribution can be regulated by adjusting the buffer pH. Using this support, we determined the cryo-EM structures of catalase (2.29 Å) and influenza haemagglutinin trimer (2.56 Å), which exhibited strong preferred orientations using a conventional cryo-vitrification protocol. We further show that the HFBI film is suitable to obtain high-resolution structures of small proteins, including aldolase (150 kDa, 3.28 Å) and haemoglobin (64 kDa, 3.6 Å). Our work suggests that HFBI films may have broad future applications in increasing the success rate and efficiency of cryo-EM.

PubMed: 34907237
DOI: 10.1038/s41467-021-27596-8

実験手法

ELECTRON MICROSCOPY (3.28 Å)