TY - JOUR A1 - Brechun, Katherine Emily A1 - Arndt, Katja Maren A1 - Woolley, G. Andrew T1 - Selection of protein-protein interactions of desired affinities with a bandpass circuit JF - Journal of molecular biology : JMB N2 - We have developed a genetic circuit in Escherichia coli that can be used to select for protein-protein interactions of different strengths by changing antibiotic concentrations in the media. The genetic circuit links protein-protein interaction strength to beta-lactamase activity while simultaneously imposing tuneable positive and negative selection pressure for beta-lactamase activity. Cells only survive if they express interacting proteins with affinities that fall within set high- and low-pass thresholds; i.e. the circuit therefore acts as a bandpass filter for protein-protein interactions. We show that the circuit can be used to recover protein-protein interactions of desired affinity from a mixed population with a range of affinities. The circuit can also be used to select for inhibitors of protein-protein interactions of defined strength. (C) 2018 Elsevier Ltd. All rights reserved. KW - synthetic biology KW - genetic circuit KW - biological engineering KW - protein-protein interactions KW - twin-arginine translocation KW - selection system Y1 - 2018 U6 - https://doi.org/10.1016/j.jmb.2018.11.011 SN - 0022-2836 SN - 1089-8638 VL - 431 IS - 2 SP - 391 EP - 400 PB - Elsevier CY - London ER - TY - GEN A1 - Dahmani, Ismail A1 - Ludwig, Kai A1 - Chiantia, Salvatore T1 - Influenza A matrix protein M1 induces lipid membrane deformation via protein multimerization T2 - Postprints der Universität Potsdam Mathematisch-Naturwissenschaftliche Reihe N2 - The matrix protein M1 of the Influenza A virus (IAV) is supposed to mediate viral assembly and budding at the plasma membrane (PM) of infected cells. In order for a new viral particle to form, the PM lipid bilayer has to bend into a vesicle toward the extracellular side. Studies in cellular models have proposed that different viral proteins might be responsible for inducing membrane curvature in this context (including M1), but a clear consensus has not been reached. In the present study, we use a combination of fluorescence microscopy, cryogenic transmission electron microscopy (cryo-TEM), cryo-electron tomography (cryo-ET) and scanning fluorescence correlation spectroscopy (sFCS) to investigate M1-induced membrane deformation in biophysical models of the PM. Our results indicate that M1 is indeed able to cause membrane curvature in lipid bilayers containing negatively charged lipids, in the absence of other viral components. Furthermore, we prove that protein binding is not sufficient to induce membrane restructuring. Rather, it appears that stable M1–M1 interactions and multimer formation are required in order to alter the bilayer three-dimensional structure, through the formation of a protein scaffold. Finally, our results suggest that, in a physiological context,M1-induced membrane deformation might be modulated by the initial bilayer curvature and the lateral organization of membrane components (i.e. the presence of lipid domains). T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 768 KW - confocal microscopy KW - influenza KW - lipid membranes KW - membranes KW - protein-protein interactions KW - viral matrix proteins Y1 - 2019 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-438689 SN - 1866-8372 IS - 768 ER - TY - JOUR A1 - Dahmani, Ismail A1 - Ludwig, Kai A1 - Chiantia, Salvatore T1 - Influenza A matrix protein M1 induces lipid membrane deformation via protein multimerization JF - Bioscience Reports N2 - The matrix protein M1 of the Influenza A virus (IAV) is supposed to mediate viral assembly and budding at the plasma membrane (PM) of infected cells. In order for a new viral particle to form, the PM lipid bilayer has to bend into a vesicle toward the extracellular side. Studies in cellular models have proposed that different viral proteins might be responsible for inducing membrane curvature in this context (including M1), but a clear consensus has not been reached. In the present study, we use a combination of fluorescence microscopy, cryogenic transmission electron microscopy (cryo-TEM), cryo-electron tomography (cryo-ET) and scanning fluorescence correlation spectroscopy (sFCS) to investigate M1-induced membrane deformation in biophysical models of the PM. Our results indicate that M1 is indeed able to cause membrane curvature in lipid bilayers containing negatively charged lipids, in the absence of other viral components. Furthermore, we prove that protein binding is not sufficient to induce membrane restructuring. Rather, it appears that stable M1–M1 interactions and multimer formation are required in order to alter the bilayer three-dimensional structure, through the formation of a protein scaffold. Finally, our results suggest that, in a physiological context,M1-induced membrane deformation might be modulated by the initial bilayer curvature and the lateral organization of membrane components (i.e. the presence of lipid domains). KW - confocal microscopy KW - influenza KW - lipid membranes KW - membranes KW - protein-protein interactions KW - viral matrix proteins Y1 - 2019 U6 - https://doi.org/10.1042/BSR20191024 SN - 0144-8463 SN - 1573-4935 VL - 39 IS - 8 PB - Portland Press CY - Colchester ER - TY - JOUR A1 - Federico, Stefania A1 - Pierce, Benjamin F. A1 - Piluso, Susanna A1 - Wischke, Christian A1 - Lendlein, Andreas A1 - Neffe, Axel T. T1 - Design of Decorin-Based Peptides That Bind to CollagenI and their Potential as Adhesion Moieties in Biomaterials JF - Angewandte Chemie : a journal of the Gesellschaft Deutscher Chemiker ; International edition N2 - Mimicking the binding epitopes of protein-protein interactions by using small peptides is important for generating modular biomimetic systems. A strategy is described for the design of such bioactive peptides without accessible structural data for the targeted interaction, and the effect of incorporating such adhesion peptides in complex biomaterial systems is demonstrated. The highly repetitive structure of decorin was analyzed to identify peptides that are representative of the inner and outer surface, and it was shown that only peptides based on the inner surface of decorin bind to collagen. The peptide with the highest binding affinity for collagenI, LHERHLNNN, served to slow down the diffusion of a conjugated dye in a collagen gel, while its dimer could physically crosslink collagen, thereby enhancing the elastic modulus of the gel by one order of magnitude. These results show the potential of the identified peptides for the design of biomaterials for applications in regenerative medicine. KW - biomaterials KW - collagen KW - gels KW - peptides KW - protein-protein interactions Y1 - 2015 U6 - https://doi.org/10.1002/anie.201505227 SN - 1433-7851 SN - 1521-3773 VL - 54 IS - 37 SP - 10980 EP - 10984 PB - Wiley-VCH CY - Weinheim ER -