Expression, Purification, and Structural Characterization of Membrane Proteins: A Case Study of the ABC Transporter MsbA.

Hanieh Bahramimoghaddam, Arthur Laganowsky

Abstract read

In one paragraph

Article in Methods in molecular biology (Clifton, N.J.), 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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1 · What the graph read from it

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0 citing papers in PubMed.

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4 · The record

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5 · Who and what money

Authors and funding

2 authors.

Hanieh BahramimoghaddamDepartment of Chemistry, Texas A&M University, College Station, TX, USA.

Arthur LaganowskyDepartment of Chemistry, Texas A&M University, College Station, TX, USA. alaganowsky@chem.tamu.edu.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Integral membrane proteins play essential roles in cellular physiology but present formidable challenges in their overexpression, purification, and structural characterization. This chapter outlines a detailed workflow for the expression, purification, and structural analysis of the ABC transporter MsbA from Pseudomonas aeruginosa (PaMsbA). This system exemplifies how biochemical optimization, native mass spectrometry (MS), and cryo-electron microscopy (cryo-EM) can be integrated to investigate membrane protein function and structure. The workflow includes expression and purification of PaMsbA from Escherichia coli for structural and biophysical studies. A unique feature of PaMsbA is its activation by divalent metals (Zn²⁺, Ni²⁺, and Mn²⁺), which is necessary for vanadate trapping and high-resolution cryo-EM structure determination. Together, these approaches establish a framework for preparing high-quality membrane protein samples for mechanistic and structural investigations.

Indexed as

ATP-Binding Cassette TransportersBacterial ProteinsMembrane ProteinsPseudomonas aeruginosaCloning, MolecularCryoelectron MicroscopyEscherichia coliGene ExpressionMass SpectrometryProtein ConformationRecombinant ProteinsATP-Binding Cassette TransportersBacterial ProteinsMembrane ProteinsMsbA protein, BacteriaRecombinant ProteinsABC transporterCryo-electron microscopyDivalent metalsLipid bindingMembrane proteinMsbANative mass spectrometryProtein purificationPseudomonas aeruginosaStructural biology

Identifiers

PMID42681345

DOI10.1038/nature13419

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.