Single particle cryo-EM reconstruction of 52 kDa streptavidin at 3.2 Angstrom resolution

被引：125

作者：

Fan, Xiao ^{[1
,2
]}

Wang, Jia ^{[1
]}

Zhang, Xing ^{[1
]}

Yang, Zi ^{[1
,2
]}

Zhang, Jin-Can ^{[3
]}

Zhao, Lingyun ^{[1
]}

Peng, Hai-Lin ^{[3
]}

Lei, Jianlin ^{[1
,2
]}

Wang, Hong-Wei ^{[1
,2
]}

机构：

[1] Tsinghua Univ, Beijing Adv Innovat Ctr Struct Biol, Beijing Frontier Res Ctr Biol Struct, Minist Educ,Key Lab Prot Sci,Sch Life Sci, Beijing 100084, Peoples R China

[2] Tsinghua Univ, Tsinghua Peking Joint Ctr Life Sci, Beijing 100084, Peoples R China

[3] Peking Univ, Acad Adv Interdisciplinary Studies, Beijing 100871, Peoples R China

来源：

NATURE COMMUNICATIONS | 2019年 / 10卷 / 1期

基金：

中国国家自然科学基金;

关键词：

BEAM-INDUCED MOTION; PHASE-PLATE; FOCUSED CLASSIFICATION; MICROSCOPY; ORIENTATION; ADSORPTION; GRAPHENE;

D O I：

10.1038/s41467-019-10368-w

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

The fast development of single-particle cryogenic electron microscopy (cryo-EM) has made it more feasible to obtain the 3D structure of well-behaved macromolecules with a molecular weight higher than 300 kDa at -3 angstrom resolution. However, it remains a challenge to obtain the high-resolution structures of molecules smaller than 200 kDa using single-particle cryo-EM. In this work, we apply the Cs-corrector-VPP-coupled cryo-EM to study the 52 kDa streptavidin (SA) protein supported on a thin layer of graphene and embedded in vitreous ice. We are able to solve both the apo-SA and biotin-bound SA structures at near-atomic resolution using single-particle cryo-EM. We demonstrate that the method has the potential to determine the structures of molecules as small as 39 kDa.

引用

页数：11

共 56 条

[1]

Adams P.D., 2010, PHENIX: a comprehensive Python-based system for macromolecular structure solution

[2] Sampling the conformational space of the catalytic subunit of human γ-secretase [J].