Open AccessA ‘Build and Retrieve’ methodology to simultaneously solve cryo-EM structures of membrane proteins
Author(s) -
ChihChia Su,
Meinan Lyu,
Christopher E. Morgan,
Jani Reddy Bolla,
Carol V. Robinson,
Edward Yu
Publication year - 2021
Publication title -
nature methods
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 19.469
H-Index - 318
eISSN - 1548-7105
pISSN - 1548-7091
DOI - 10.1038/s41592-020-01021-2
Subject(s) - cryo electron microscopy , structural biology , membrane protein , proteomics , resolution (logic) , lysis , membrane , biological system , computational biology , biophysics , sample preparation , biology , chemistry , nanotechnology , materials science , computer science , biochemistry , chromatography , artificial intelligence , gene
Single-particle cryo-electron microscopy (cryo-EM) has become a powerful technique in the field of structural biology. However, the inability to reliably produce pure, homogeneous membrane protein samples hampers the progress of their structural determination. Here, we develop a bottom-up iterative method, Build and Retrieve (BaR), that enables the identification and determination of cryo-EM structures of a variety of inner and outer membrane proteins, including membrane protein complexes of different sizes and dimensions, from a heterogeneous, impure protein sample. We also use the BaR methodology to elucidate structural information from Escherichia coli K12 crude membrane and raw lysate. The findings demonstrate that it is possible to solve high-resolution structures of a number of relatively small (<100 kDa) and less abundant (<10%) unidentified membrane proteins within a single, heterogeneous sample. Importantly, these results highlight the potential of cryo-EM for systems structural proteomics.