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Co-assembly, spatiotemporal control and morphogenesis of a hybrid protein–peptide system

Abstract

Controlling molecular interactions between bioinspired molecules can enable the development of new materials with higher complexity and innovative properties. Here we report on a dynamic system that emerges from the conformational modification of an elastin-like protein by peptide amphiphiles and with the capacity to access, and be maintained in, non-equilibrium for substantial periods of time. The system enables the formation of a robust membrane that displays controlled assembly and disassembly capabilities, adhesion and sealing to surfaces, self-healing and the capability to undergo morphogenesis into tubular structures with high spatiotemporal control. We use advanced microscopy along with turbidity and spectroscopic measurements to investigate the mechanism of assembly and its relation to the distinctive membrane architecture and the resulting dynamic properties. Using cell-culture experiments with endothelial and adipose-derived stem cells, we demonstrate the potential of this system to generate complex bioactive scaffolds for applications such as tissue engineering.

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Figure 1: Self-assembly, dynamic properties and architecture of the hybrid ELP5/PAK3 system.
Figure 2: Molecular interactions and composition of the ELP5/PAK3 membrane.
Figure 3: Relation between the molecular mechanism, membrane architecture and properties of the ELP5/PAK3 system.
Figure 4: Application in tissue engineering.

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Acknowledgements

The work was supported by the European Research Council Starting Grant (STROFUNSCAFF), the European Commission under FP7 and H2020 programs ((NMP3-LA-2011-263363, HEALTH-F4-2011-278557, PITN-GA-2012-317304, MSCA-ITN-2014-ETN- 642687, 642687 H2020-NMP-2014-646075), the Ministry of Economy and Competitiveness (Spain) (MAT2012-38043-C02-01, MAT2013-41723-R, MAT2013-42473-R) the Junta de Castilla y Leon (VA244U13, VA313U14) and the Portuguese Foundation for Science and Technology, grants PTDC/EBB-BIO/114523/2009 and SFRH/BD/44977/2008. Additional support was obtained from the Bilateral Program Portugal–Spain Integrated Actions 2011 (E-50/11) and Marie Curie Career Integration Grant 618335. The authors thank the European Synchrotron Research Facility for access to synchrotron beamline BM29 and P. Pernot for support during the experiments, and C. López (Centres Científics i Tecnològics University of Barcelona), C. Semino (Institut Químic de Sarrià), E. Rebollo (Advanced Fluorescence Microscopy Unit in the Molecular Biology Institute of Barcelona), J. P. Aguilar, R. Doodkorte, A. Amzour and the technical staff of the Material Characterization Laboratory and Nanovision Laboratory at the Queen Mary University of London for the constructive discussions and contributions in this study.

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A.M., K.H.S. and K.E.I. conceived the project. D.S.F. and H.S.A. performed the synthesis and chemical characterization of some PAs used in this study. E.C. carried out cell-culture studies. O.S and R.B. collected and analysed SAXS data. R.P.R. performed and analysed the TOF-SIM experiments and K.E.I. performed experiments. M.A., J.C.R. and R.L.R. provided facilities to perform experiments. A.M., K.E.I., A.M.-M., H.S.A., L.B., R.B. and F.S. interpreted the data and wrote the manuscript. All the authors discussed the results and commented on the manuscript.

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Correspondence to Alvaro Mata.

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Inostroza-Brito, K., Collin, E., Siton-Mendelson, O. et al. Co-assembly, spatiotemporal control and morphogenesis of a hybrid protein–peptide system. Nature Chem 7, 897–904 (2015). https://doi.org/10.1038/nchem.2349

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