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

• Published in: Nanotechnology Vol. 25; no. 28; pp. 285101 - 7
• Main Authors: Paulin, Sarah, Jamshad, Mohammed, Dafforn, Timothy R, Garcia-Lara, Jorge, Foster, Simon J, Galley, Nicola F, Roper, David I, Rosado, Helena, Taylor, Peter W
• Format: Journal Article
• Language: English
• Published: Bristol IOP Publishing 18.07.2014; Institute of Physics
• Subjects: antibiotic resistance; Bacteria; Bacterial Proteins - chemistry; Biological and medical sciences; Biotechnology; Catechin - analogs & derivatives; Catechin - chemistry; Cell Division - physiology; Cross-disciplinary physics: materials science; rheology; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; Gallates; immunoaffinity chromatography; Lipid Bilayers - chemistry; lipid nanoparticles; Maleates - chemistry; Materials science; Membrane Proteins - chemistry; Membranes; Methods. Procedures. Technologies; Molecular structure; Nanocrystalline materials; Nanoscale materials and structures: fabrication and characterization; Others; Penicillin-Binding Proteins - chemistry; Peptide Synthases - chemistry; Physics; poly(styrene-co-maleic acid); Polystyrenes - chemistry; Proteins; Purification; Shape memory alloys; Staphylococcus aureus - chemistry; Surface-Active Agents - chemistry; Various methods and equipments; Drug; Structure function; Membrane protein; Purification; Nanoparticle; Lactam; Lipids; Polymer; Surfactant; Lipid bilayers; Binding protein; Macromolecule; Maleic acid; Bacteria; Disruption; Nanostructured materials
• ISSN: 0957-4484; 1361-6528
• DOI: 10.1088/0957-4484/25/28/285101

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.