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Open AccessArticle

A Versatile and Robust Serine Protease Inhibitor Scaffold from Actinia tenebrosa

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Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, QLD 4059, Australia
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School of Biomedical Sciences, Faculty of Health, Queensland University of Technology, Brisbane, QLD 4000, Australia
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School of Earth, Environmental and Biological Sciences, Science and Engineering Faculty, Queensland University of Technology, Brisbane, QLD 4000, Australia
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Institute for Future Environments, Queensland University of Technology, Brisbane, QLD 4000, Australia
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Institute for Molecular Bioscience, University of Queensland, St Lucia, QLD 4072, Australia
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Centre for Advanced Imaging, University of Queensland, St Lucia, QLD 4072, Australia
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Centre for Biodiversity Dynamics, Department of Biology, Norwegian University of Science and Technology, 7491 Trondheim, Norway
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Centre for Ecological and Evolutionary Synthesis, Department of Biosciences, University of Oslo, PO Box 1066 Blindern, 0316 Oslo, Norway
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Department of Medical Biosciences, Umeå University, 901 87 Umeå, Sweden
*
Author to whom correspondence should be addressed.
Mar. Drugs 2019, 17(12), 701; https://doi.org/10.3390/md17120701
Received: 29 October 2019 / Revised: 10 December 2019 / Accepted: 10 December 2019 / Published: 12 December 2019
Serine proteases play pivotal roles in normal physiology and a spectrum of patho-physiological processes. Accordingly, there is considerable interest in the discovery and design of potent serine protease inhibitors for therapeutic applications. This led to concerted efforts to discover versatile and robust molecular scaffolds for inhibitor design. This investigation is a bioprospecting study that aims to isolate and identify protease inhibitors from the cnidarian Actinia tenebrosa. The study isolated two Kunitz-type protease inhibitors with very similar sequences but quite divergent inhibitory potencies when assayed against bovine trypsin, chymostrypsin, and a selection of human sequence-related peptidases. Homology modeling and molecular dynamics simulations of these inhibitors in complex with their targets were carried out and, collectively, these methodologies enabled the definition of a versatile scaffold for inhibitor design. Thermal denaturation studies showed that the inhibitors were remarkably robust. To gain a fine-grained map of the residues responsible for this stability, we conducted in silico alanine scanning and quantified individual residue contributions to the inhibitor’s stability. Sequences of these inhibitors were then used to search for Kunitz homologs in an A. tenebrosa transcriptome library, resulting in the discovery of a further 14 related sequences. Consensus analysis of these variants identified a rich molecular diversity of Kunitz domains and expanded the palette of potential residue substitutions for rational inhibitor design using this domain. View Full-Text
Keywords: Actinia tenebrosa; kallikrein-related peptidases; Kunitz inhibitor; serine protease; mass spectrometry imaging; molecular dynamics simulation Actinia tenebrosa; kallikrein-related peptidases; Kunitz inhibitor; serine protease; mass spectrometry imaging; molecular dynamics simulation
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Chen, X.; Leahy, D.; Van Haeften, J.; Hartfield, P.; Prentis, P.J.; van der Burg, C.A.; Surm, J.M.; Pavasovic, A.; Madio, B.; Hamilton, B.R.; King, G.F.; Undheim, E.A.B.; Brattsand, M.; Harris, J.M. A Versatile and Robust Serine Protease Inhibitor Scaffold from Actinia tenebrosa. Mar. Drugs 2019, 17, 701.

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