Subtraction of liposome signals in cryo-EM structural determination of protein-liposome complexes

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外文摘要：Reconstituting membrane proteins in liposomes and determining their structure is a common method for determining membrane protein structures using single-particle cryo-electron microscopy(cryo-EM).However,the strong signal of li-posomes under cryo-EM imaging conditions often interferes with the structural determination of the embedded membrane proteins.Here,we propose a liposome signal subtraction method based on single-particle two-dimensional(2D)classifica-tion average images,aimed at enhancing the reconstruction resolution of membrane proteins.We analyzed the signal distri-bution characteristics of liposomes and proteins within the 2D classification average images of protein-liposome complexes in the frequency domain.Based on this analysis,we designed a method to subtract the liposome signals from the original particle images.After the subtraction,the accuracy of single-particle three-dimensional(3D)alignment was improved,enhancing the resolution of the final 3D reconstruction.We demonstrated this method using a PIEZO 1-proteoliposome dataset by improving the resolution of the PIEZO 1 protein.

外文关键词：

cryo-EMprotein-liposome complexesliposome signal subtraction2D classification averaging

作者：

李首卿、李明、王玉梅、李雪明

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作者单位：

Institute of Physics,Chinese Academy of Sciences,Beijing 100190,China

University of Chinese Academy of Sciences,Beijing 100049,China

Beijing Branch of Songshan Lake Materials Laboratory,Beijing 100190,China

Key Laboratory for Protein Sciences of Ministry of Education,School of Life Sciences,Tsinghua University,Beijing 100084,China

State Key Laboratory of Membrane Biology,School of Life Sciences,Tsinghua University,Beijing 100084,China

Tsinghua-Peking Joint Center for Life Sciences,Beijing 100084,China

Beijing Frontier Research Center for Biological Structure,Beijing 100084,China

School of Life Sciences,Tsinghua University,Beijing 100084,China

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基金：

National Natural Science Foundation of ChinaNational Natural Science Foundation of ChinaFund from the Tsinghua-Peking Joint Center for Life sciencesBeijing Frontier Research Center for Biological Structure

项目编号：

3224102392254306

出版年：

2024

DOI：

10.1088/1674-1056/ad4cdb

中国物理B(英文版)

中国物理学会和中国科学院物理研究所

中国物理B(英文版)

CSTPCDEI

影响因子：0.995

ISSN：1674-1056

年,卷(期)：2024.33(8)