2024Li Subtraction

Citation

S. Li, M. Li, Y. Wang, and X. Li, “Subtraction of liposome signals in cryo-EM structural determination of protein-liposome complexes,” Chinese Physics B, 2024.

Abstract

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 liposomes 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) classification average images, aimed at enhancing the reconstruction resolution of membrane proteins. We analyzed the signal distribution 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 PIEZO1-proteoliposome dataset by improving the resolution of the PIEZO1 protein.

Links

https://iopscience.iop.org/article/10.1088/1674-1056/ad4cdb/meta