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Research

Bacteria at interfaces

We study the interactions of bacterial cells with polymer surfaces of varying physico-chemical characteristics to understand and analyse the strategy of bacterial adsorption, adhesion, and biofilm development. With this approach we hope to identify surface properties that can be exploited to prevent extensive spreading of pathogens. We are exploiting the adhesion machinery of bacterial cells to build surface/peptide libraries for the detection of pathogenic strains. Polymer platforms are used as substrates for the immobilization of peptides and bacteria. We will develop a complex biofilm in vitro model to study the interactions between biofilm microbiota and eukaryotic cells. It will enable the investigation of complex developmental stages of biofilms and reveal the unique bacteria-host interactions at the cellular and the molecular level.

Different techniques to characterize bacterial interactions on surfaces with varying properties.

Bacteria meet Microfabs

The full extend of how bacteria organize biomolecules is not understood. We study the interactions of bacterial biomolecules with membranes using a range of in vivo and in vitro assays. We are developing microchamber designs to analyse protein and lipid localisation in vivo. We are using a range of biophysical techniques to analyse protein-lipid interactions in vitro. One of our aims is to develop cell free expression platforms to characterize the behavior of shape-determining and cell division proteins in liposome environments - in collaboration with Dr. Toshihisa Osaki (The University of Tokyo) and Dr. Piotr Garstecki (Polish Academy of Sciences).

Bacterial Morphology/Growth

We are interested to understand how bacteria cells control their geometry and how bacterial cells are responding to morphological changes. We employ different microchamber designs to morph bacterial cells and track the morphological recovery.

Bacteria can be reshaped and the localization of biomolecules is studied in dependence of curvature. (A) Confinement of spheroplasts from E. coli in microchambers of different curvatures (B) stained with NAO (red) and DAPI (blue) for cardiolipin domains and DNA, respectively (Renner and Weibel, 2011, PNAS). (C) Deformed bacteria cells (E. coli) confined in angular microchambers responding to external force and (D) filaments stained for DNA (Renner et al., 2013, PLoS One).
Bacterial growth in agarose microchambers: Cells were filamented by infusing the agarose with 25 µg/mL cephalexin. Cells readily adapt microchamber topography. Scale bar: 20 µm.
Projection of short MreB filaments in filamented and bent Escherichia coli cell in angled microchamber. (Renner et. al, 2013, PLoS One).

Biomimetic Lipid-Bilayer on Polymer Cushions

We are using supported lipid bilayer systems to mimic biological membranes on different substrates (silicon dioxide, glass, polymer substrates) to understand protein-lipid interactions of membrane-bound and transmembrane proteins (prokaryotic and eukaryotic proteins).

Versatile biocompatible polymer platform. Maleic anhydride copolymers with varying physico-chemical properties were used as polymer cushions, spanning hydrophilic to hydrophobic characteristics. The maleic anhydride group hydrolyses in water and can be used to covalently immobilize biomolecules. (Macromol Biosci, 2005, 5, 890-895; Express Polymer Letters, 2009, 3, 33-38)

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Supported lipid bilayer can be formed on a variety of substrates; solid, polymer-supported or pore free-standing. We formed lipid bilayers on maleic anhydride polymers as cushions to incorporate functional transmembrane proteins. We successfully integrated β-amyloid cleaving enzyme. As each cushion exhibits different physico-chemical properties, the resulting behavior of the lipid bilayer and transmembrane protein could be exactly adjusted to the specific environmental requirements. (J Phys Chem B, 2008, 112, 6373-6378; Biointerphases, 2009, 4, 1-6; Soft Matter, 2010, 6, 5382-5389; Soft Matter, 2010, 6, 937-944)

Protein-Lipid Interactions

The prokaryotic division oscillator MinD showed strong interactions with cardiolipin microdomains in localization studies (PNAS, 2011, 108, 6264-6269). We analysed the membrane binding affinity, kinetics and ATPase activity of MinD on supported lipid bilayers of varying compositions. Increased concentrations of anionic phospholipids (CL, PG) increase the binding affinity and decreased the ATPase activity of MinD (JBC, 2012, 287, 38835-38844).