Membrane protein structure; Novel Antibiotics; Membrane protein function; Bacterial outer membrane; BamA; Omp85 insertase; Nuclear Magnetic Resonance (NMR)
Kaur Hundeep, Jakob Roman P., Marzinek Jan K., Green Robert, Imai Yu, Bolla Jani Reddy, Agustoni Elia, Robinson Carol V., Bond Peter J., Lewis Kim, Maier Timm, Hiller Sebastian (2021), The antibiotic darobactin mimics a β-strand to inhibit outer membrane insertase, in Nature
, 593(7857), 125-129.
Mas Guillaume, Burmann Björn M., Sharpe Timothy, Claudi Beatrice, Bumann Dirk, Hiller Sebastian (2020), Regulation of chaperone function by coupled folding and oligomerization, in Science Advances
, 6(43), eabc5822.
Kaur Hundeep, Grahl Anne, Hartmann Jean-Baptiste, Hiller Sebastian (2020), Expression, Purification, and Structural Biology of Membrane Proteins, in Methods Molecular Biology
Rath Parthasarathi, Sharpe Timothy, Hiller Sebastian (2020), The electrostatic core of the outer membrane protein X from E. coli, in Biochimica et Biophysica Acta (BBA) - Biomembranes
, 1862(1), 183031-183031.
Luther Anatol, Urfer Matthias, Zahn Michael, Müller Maik, Wang Shuang-Yan, Mondal Milon, Vitale Alessandra, Hartmann Jean-Baptiste, Sharpe Timothy, Monte Fabio Lo, Kocherla Harsha, Cline Elizabeth, Pessi Gabriella, Rath Parthasarathi, Modaresi Seyed Majed, Chiquet Petra, Stiegeler Sarah, Verbree Carolin, Remus Tobias, Schmitt Michel, Kolopp Caroline, Westwood Marie-Anne, Desjonquères Nicolas, Brabet Emile, et al. (2019), Chimeric peptidomimetic antibiotics against Gram-negative bacteria, in Nature
, 576(7787), 452-458.
Imai Yu, Meyer Kirsten J., Iinishi Akira, Favre-Godal Quentin, Green Robert, Manuse Sylvie, Caboni Mariaelena, Mori Miho, Niles Samantha, Ghiglieri Meghan, Honrao Chandrashekhar, Ma Xiaoyu, Guo Jason J., Makriyannis Alexandros, Linares-Otoya Luis, Böhringer Nils, Wuisan Zerlina G., Kaur Hundeep, Wu Runrun, Mateus André, Typas Athanasios, Savitski Mikhail M., Espinoza Josh L., O’Rourke Aubrie, et al. (2019), A new antibiotic selectively kills Gram-negative pathogens, in Nature
, 576(7787), 459-464.
Kaur Hundeep, Hartmann Jean-Baptiste, Jakob Roman P., Zahn Michael, Zimmermann Iwan, Maier Timm, Seeger Markus A., Hiller Sebastian (2019), Identification of conformation-selective nanobodies against the membrane protein insertase BamA by an integrated structural biology approach, in Journal of Biomolecular NMR
, 73(6-7), 375-384.
Bibow Stefan, Hiller Sebastian (2019), A guide to quantifying membrane protein dynamics in lipids and other native‐like environments by solution‐state NMR spectroscopy, in The FEBS Journal
, 286(9), 1610-1623.
Rath Parthasarathi, Sharpe Timothy, Kohl Bastian, Hiller Sebastian (2019), Two-State Folding of the Outer Membrane Protein X into a Lipid Bilayer Membrane, in Angewandte Chemie International Edition
Hartmann Jean-Baptiste, Zahn Michael, Burmann Irena Matečko, Bibow Stefan, Hiller Sebastian (2018), Sequence-Specific Solution NMR Assignments of the β-Barrel Insertase BamA to Monitor Its Conformational Ensemble at the Atomic Level, in Journal of the American Chemical Society
, 140, 11252.
In recent years, multidrug resistance (MDR) of microorganisms against antibiotics is worldwide on the rise. The currently most threatening species include the Gram-negative Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp. In these Gram-negative bacteria, the outer membrane presumably provides an ideal target for novel antibiotics. Among the typically around 80 outer membrane proteins (Omps), only two - BamA and LptD - are generally essential and highly conserved across Gram-negative bacteria. LptD is efficiently targeted by the protein epitope mimic (PEM) POL7080, a cyclic peptide with potent antimicrobial activity against P. aeruginosa that is currently in clinical trials (Polyphor AG, Allschwil BL). With increased spreading of MDR-strains, the identification of additional new targets is urgently needed.BamA is an integral membrane protein, forming the central unit of the Bam complex. Deletion of BamA and even certain single-point mutations are lethal to E. coli, underscoring the relevance as an outer membrane antibiotic target. BamA inserts a broad range of different substrates into the membrane, but despite knowledge of the structure, the functional mechanism of BamA has so far remained unclear. Since the interaction with its substrates is only semi-specific, there is a high potential for specific high-affinity binders that may block BamA function. A description of the BamA functional mechanism at the atomic level would thus provide essential insights, how such new antibiotics may be developed. In collaboration with our industry partner Polyphor AG (Allschwil BL), we want to investigate the mechanism of action of a new family of PEM antibiotics that have been shown to be highly active against all Gram-negative ESKAPE pathogens. The new family of PEMs is presumably targeting other members of the ß-barrel outer membrane protein family such as the insertase BamA.We will pursue the following objectives:1.) An atomic level description of the functional mechanism of substrate insertion by BamA.2.) An atomic level structural and functional description of its inhibition by PEM antibiotics and newly identified natural inhibitors.3.) Establishment of a quantitative assay suitable for high-throughput screening of new BamA inhibitors.To achieve these goals, we will use solution NMR spectroscopy, as well as X-ray crystallography, and biochemical and biophysical methods. We base our work on strong expertise in these techniques, as well as on BamA and Bam complex biochemistry.