Poster Presentation 2014 International Biophysics Congress

The structure of lipopolysaccharide from rough strains of Escherichia coli at the air – liquid interface (#458)

Anton P. Le Brun 1 , Luke A. Clifton 2 , Cnadice E. Halbert 3 , Binhua Lin 4 , Mati Meron 4 , Peter J. Holden 1 , Jeremy H. Lakey 5 , Stephen A. Holt 1
  1. Australian Nuclear Science Technology Organisation, Lucas Heights, NSW, Australia
  2. ISIS Neutron Facility, STFC Rutherford Appleton Laboratory, Didcot, Oxfordshire, UK
  3. Spallation Neutron Source, Oak Ridge National Laboratory, Oak Ridge, TN, USA
  4. CARS, University of Chicago, Chicago, IL, USA
  5. Institute for Cell and Molecular Biosciences, University of Newcastle upon Tyne, Newcastle upon Tyne, UK

The outer membrane (OM) of the Gram-negative bacterial cell envelope is an important biological interface as it initiates bacterial cell adhesion, activates innate immunity during infection, withstands environmental stresses and acts as a gate keeper to nutrients and toxins. The OM is a complex structure consisting of an asymmetric bilayer with an inner leaflet of phospholipids and an outer leaflet of lipopolysaccharide (LPS) with LPS consisting of approximately 75 % of the Gram-negative bacterial surface. Due to the complexity of LPS there are very few OM models that include this important molecule and thus little is known about LPS structure at interfaces. In this work we used a truncated LPS from E. coli (Rc-LPS, consisting of Lipid A and the first seven sugars of the core polysaccharide) to form stable monolayers at the air – liquid interface. Neutron and X-ray reflectometry were used to probe the vertical structure of the monolayers, while the lateral structure was studied with grazing incidence X-ray diffraction and Brewster angle microscopy. It was found that at surface pressures above 20 mN/m the RcLPS monolayers could be resolved as hydrocarbon tails, inner headgroup, and outer headgroup of polysaccharide with increasing solvation from tails to outer headgroup. The lateral organisation of the hydrocarbon tails was of an oblique hexagonal unit cell across all surface pressures with only the tilt angle of the chains changing with surface pressure [1]. The study of the monolayer structure provides the first complete analysis of a realistic E. coli OM surface model.