Dynamic covalent chemistry enables formation of antimicrobial peptide quaternary assemblies in a completely abiotic manner

Abstract

Naturally occurring peptides and proteins often use dynamic disulfide bonds to impart defined tertiary/quaternary structures for the formation of binding pockets with uniform size and function. Although peptide synthesis and modification are well established, controlling quaternary structure formation remains a significant challenge. Here, we report the facile incorporation of aryl aldehyde and acyl hydrazide functionalities into peptide oligomers via solid-phase copper-catalysed azide–alkyne cycloaddition (SP-CuAAC) click reactions. When mixed, these complementary functional groups rapidly react in aqueous media at neutral pH to form peptide–peptide intermolecular macrocycles with highly tunable ring sizes. Moreover, sequence-specific figure-of-eight, dumbbell-shaped, zipper-like and multi-loop quaternary structures were formed selectively. Controlling the proportions of reacting peptides with mismatched numbers of complementary reactive groups results in the formation of higher-molecular-weight sequence-defined ladder polymers. This also amplified antimicrobial effectiveness in select cases. This strategy represents a general approach to the creation of complex abiotic peptide quaternary structures.

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Figure 1: Synthesis of DC–peptides.
Figure 2: Formation of DC–peptide quaternary structures.
Figure 3: Tricine SDS–PAGE of DC–peptide assemblies.
Figure 4: Vernier templated DC–peptide ladder polymerization.
Figure 5: Hydrodynamic radii of DC–peptides and quaternary assemblies.
Figure 6: Antimicrobial activity of DC–peptides and assemblies against the Gram-positive bacterium S. aureus.

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Acknowledgements

Financial support for the presented work was provided by the DARPA Fold-Fx programme (N66001-14-2-4051) and the Welch Regents Chair (F-0046). The authors thank M. Persons of the Proteomics facility at University of Texas (UT) at Austin for aid with MALDI–TOF MS acquisition, S. Sorey of the NMR facility at UT Austin for aid with 2D-DOSY-NMR acquisition and I. Riddington of the Mass Spectrometry facility at UT Austin for aid with HRMS acquisition.

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J.F.R. and E.V.A. devised the conducted experiments. J.F.R. wrote the manuscript, collected the MALDI–TOF MS data, collected NMR data and conducted SDS–PAGE experiments. J.L.D. and M.W. designed and conducted the high-throughput luminescence assay for determination of antibiotic efficiencies. J.F.R., I.V.K., D.V.U. and R.G. contributed to the synthesis and purification of all peptides and small-molecule precursors reported. E.T.H. aided in the training for peptide synthesis and HPLC purification. All authors edited the manuscript.

Corresponding authors

Correspondence to Marvin Whiteley or Eric V. Anslyn.

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The authors declare no competing financial interests.

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Reuther, J., Dees, J., Kolesnichenko, I. et al. Dynamic covalent chemistry enables formation of antimicrobial peptide quaternary assemblies in a completely abiotic manner. Nature Chem 10, 45–50 (2018). https://doi.org/10.1038/nchem.2847

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