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A 'Build and Retrieve' methodology to simultaneously solve cryo-EM structures of membrane proteins.
- Source :
-
Nature methods [Nat Methods] 2021 Jan; Vol. 18 (1), pp. 69-75. Date of Electronic Publication: 2021 Jan 06. - Publication Year :
- 2021
-
Abstract
- 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.
- Subjects :
- Bacterial Proteins metabolism
Cell Membrane chemistry
Escherichia coli metabolism
Humans
Membrane Proteins metabolism
Models, Molecular
Molecular Structure
Protein Conformation
Bacterial Proteins chemistry
Burkholderia pseudomallei metabolism
Cell Membrane metabolism
Cryoelectron Microscopy methods
Membrane Proteins chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1548-7105
- Volume :
- 18
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Nature methods
- Publication Type :
- Academic Journal
- Accession number :
- 33408407
- Full Text :
- https://doi.org/10.1038/s41592-020-01021-2