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TitleExploration of a workflow for the classification and identification of co-purified protein complexes yields new structures and multiple MSP assembly states.
Journal, issue, pagesPLoS One, Vol. 21, Issue 1, Page e0340207, Year 2026
Publish dateJan 27, 2026
AuthorsQingyang Zhang / Abhinandan Venkatesha Murthy / Carsten Mim /
PubMed AbstractNative protein complexes have garnered interest as targets for structural dissemination. Cryogenic electron microscopy (cryo-EM) with its ability to image protein mixtures is the most promising tool ...Native protein complexes have garnered interest as targets for structural dissemination. Cryogenic electron microscopy (cryo-EM) with its ability to image protein mixtures is the most promising tool to enable structural proteomics. Additionally, image processing has evolved and can deal with conformational and compositional heterogeneity. Integrative approaches, namely mass spectrometry in conjunction with cryo-EM, have made it possible to characterize and identify complex mixtures. However, this comes at a cost of generating models and interpreting mass spectra. Here we present a modified approach that builds on publicly available software. By generating maps around 4 Å and unsupervised model building we were able to identify the most abundant proteins in our sample. This sample consisted of co-purified membrane proteins in nanodiscs. We found a novel structure and unexpected nanodisc assemblies. Our maps imply a direct interaction between membrane proteins and membrane scaffolding proteins.
External linksPLoS One / PubMed:41592050 / PubMed Central
MethodsEM (single particle)
Resolution2.72 - 3.27 Å
Structure data

52450

9hwj

EMDB-52450, PDB-9hwj:
Structure of Undecaprenyl-phosphate 4-deoxy-4-formamido-L-arabinose transferase embedded in nanodisc
Method: EM (single particle) / Resolution: 3.27 Å

52451

9hwk

EMDB-52451, PDB-9hwk:
Structure of the Cytochrome o ubiquinol oxidase embedded in the nanodisc
Method: EM (single particle) / Resolution: 2.72 Å

52452

9hwl

EMDB-52452, PDB-9hwl:
Structure of the co-purified multidrug transporter subunit ACRB in nandisc
Method: EM (single particle) / Resolution: 3.27 Å

Chemicals

ChemComp-HEO:
HEME O

ChemComp-HEM:
PROTOPORPHYRIN IX CONTAINING FE

ChemComp-CU:
COPPER (II) ION

ChemComp-ZN:
Unknown entry

Source
  • escherichia coli (E. coli)
KeywordsMEMBRANE PROTEIN / Transferase / ELECTRON TRANSPORT / oxidase / TRANSPORT PROTEIN / Multidrug transporter

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