title: Surfactant-free purification of membrane protein complexes from bacteria: application to the staphylococcal penicillin-binding protein complex PBP2/PBP2a.
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.
authors: School of Pharmacy, University College London, 29-39 Brunswick Square, London WC1N 1AX, UK.
authors: Sarah
authors: S
authors: Paulin
authors: Mohammed
authors: M
authors: Jamshad
authors: Timothy R
authors: TR
authors: Dafforn
authors: Jorge
authors: J
authors: Garcia-Lara
authors: Simon J
authors: SJ
authors: Foster
authors: Nicola F
authors: NF
authors: Galley
authors: David I
authors: DI
authors: Roper
authors: Helena
authors: H
authors: Rosado
authors: Peter W
authors: PW
authors: Taylor
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry*; D002392:Catechin / Q000031:analogs & derivatives / Q000737:chemistry; D002455:Cell Division / Q000502:physiology; D008051:Lipid Bilayers / Q000737:chemistry; D008298:Maleates / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry*; D046915:Penicillin-Binding Proteins / Q000737:chemistry*; D010453:Peptide Synthases / Q000737:chemistry*; D011137:Polystyrenes / Q000737:chemistry; D013211:Staphylococcus aureus / Q000737:chemistry*; D013501:Surface-Active Agents / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D008051:Lipid Bilayers; D008298:Maleates; D008565:Membrane Proteins; D046915:Penicillin-Binding Proteins; D011137:Polystyrenes; D013501:Surface-Active Agents; C478699:penicillin-binding protein 2a, Streptococcus; C024074:styrene-maleic acid polymer; D002392:Catechin; C062669:epicatechin gallate; D010453:Peptide Synthases