Surfactant-free purification of membrane protein complexes from bacteria: application to the staphylococcal penicillin-binding protein complex PBP2/PBP2a

Sarah Paulin, Mohammed Jamshad, Timothy R Dafforn, Jorge Garcia-lara, Simon J Foster, Nicola F Galley, David I Roper, Helena Rosado, Peter W Taylor

Research output: Contribution to journalArticlepeer-review

40 Citations (Scopus)
548 Downloads (Pure)

Abstract

Surfactant-mediated removal of proteins from biomembranes invariably results in partial or complete loss of function and disassembly of multi-protein complexes. We determined the capacity of styrene-co-maleic acid (SMA) co-polymer to remove components of the cell division machinery from the membrane of drug-resistant staphylococcal cells. SMA-lipid nanoparticles solubilized FtsZ-PBP2-PBP2a complexes from intact cells, demonstrating the close physical proximity of these proteins within the lipid bilayer. Exposure of bacteria to (-)-epicatechin gallate, a polyphenolic agent that abolishes β-lactam resistance in staphylococci, disrupted the association between PBP2 and PBP2a. Thus, SMA purification provides a means to remove native integral membrane protein assemblages with minimal physical disruption and shows promise as a tool for the interrogation of molecular aspects of bacterial membrane protein structure and function.
Original languageEnglish
Article number285101
JournalNanotechnology
Volume25
Issue number28
DOIs
Publication statusPublished - 18 Jul 2014

Keywords

  • Staphylococcus aureus
  • poly(styrene-co-maleic acid)
  • lipid nanoparticles
  • antibiotic resistance
  • immunoaffinity chromatography

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