The C-terminus of panusin, a lobster β-defensin, is crucial for optimal antimicrobial activity and serum stability

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  • dc.contributor.author Bello Madruga, Roberto
  • dc.contributor.author Valle, Javier
  • dc.contributor.author Jiménez, M. Ángeles
  • dc.contributor.author Torrent Burgas, Marc
  • dc.contributor.author Montero Alejo, Vivian
  • dc.contributor.author Andreu Martínez, David
  • dc.date.accessioned 2023-10-06T06:03:35Z
  • dc.date.available 2023-10-06T06:03:35Z
  • dc.date.issued 2023
  • dc.description.abstract β-defensins are one of the most abundant and studied families of antimicrobial peptides (AMPs). Because of their selective toxicity to bacterial membranes and a broad spectrum of microbicidal action, β-defensins are regarded as potential therapeutic agents. This work focuses on a β-defensin-like AMP from the spiny lobster Panulirus argus (hereafter referred to as panusin or PaD). This AMP is structurally related to mammalian defensins via the presence of an αβ domain stabilized by disulfide bonds. Previous studies of PaD suggest that its C-terminus (Ct_PaD) contains the main structural determinants of antibacterial activity. To confirm this hypothesis, we made synthetic versions of PaD and Ct_PaD to determine the influence of the C-terminus on antimicrobial activity, cytotoxicity, proteolytic stability, and 3D structure. After successful solid-phase synthesis and folding, antibacterial assays of both peptides showed truncated Ct_PaD to be more active than native PaD, confirming the role of the C-terminus in activity and suggesting that cationic residues in that region enhance binding to negatively charged membranes. On the other hand, neither PaD nor Ct_PaD were hemolytic or cytotoxic in human cells. Proteolysis in human serum was also studied, showing high (>24 h) t1/2 values for PaD and lower but still considerable for Ct_PaD, indicating that the missing native disulfide bond in Ct_PaD alters protease resistance, albeit not decisively. NMR-2D experiments in water agree with the results obtained by circular dichroism (CD), where in SDS micelles, CD showed both peptides adopting an increasingly ordered structure in a hydrophobic environment, in tune with their ability to perturb bacterial membrane systems. In conclusion, while the β-defensin features of PaD are confirmed as advantageous in terms of antimicrobial activity, toxicity, and protease stability, the results of the present work suggest that these same features are preserved, even enhanced, in the structurally simpler Ct_PaD, which must therefore be viewed as a valuable lead for the development of novel anti-infectives.
  • dc.description.sponsorship Grants from MCIN, Spanish Ministry of Science and Innovation (PID2020-113184RB-C22/AEI/10.13039/501100011033 to D.A.; PID2020-114627RB-I00/AEI/10.13039/501100011033 and PDC2021-121544-I00/AEI/10.13039/501100011033 to M.T.; and PID2020-112821GB-I00/AEI/10.13039/501100011033 to M.A.J.) are acknowledged. Work at UPF also supported by La Caixa Health Foundation (grant HR17-00409 to D.A.) and by MCIN “María de Maeztu” Program for Units of Excellence in R&D (CEX2018-000792-M)) to the UPF Department of Medicine and Life Sciences at (MELIS-UPF). Work at UAB further supported by a 2022 grant from the European Society of Clinical Microbiology and Infectious Diseases (ESCMID). NMR experiments performed at the Manuel Rico NMR laboratory (LMR) of the Spanish National Research Council (CSIC), a node of the Spanish Large-Scale National Facility (ICTS R-LRB). R.B.M. is recipient of a pre-doctoral FPI scholarship (PRE2021-097678) from the Spanish Ministerio de Ciencia e Innovación.
  • dc.format.mimetype application/pdf
  • dc.identifier.citation Bello-Madruga R, Valle J, Jiménez MÁ, Torrent M, Montero-Alejo V, Andreu D. The C-terminus of panusin, a lobster β-defensin, is crucial for optimal antimicrobial activity and serum stability. Pharmaceutics. 2023 Jun 20;15(6):1777. DOI: 10.3390/pharmaceutics15061777
  • dc.identifier.doi http://dx.doi.org/10.3390/pharmaceutics15061777
  • dc.identifier.issn 1999-4923
  • dc.identifier.uri http://hdl.handle.net/10230/58053
  • dc.language.iso eng
  • dc.publisher MDPI
  • dc.relation.ispartof Pharmaceutics. 2023 Jun 20;15(6):1777
  • dc.relation.projectID info:eu-repo/grantAgreement/ES/2PE/PID2020-113184RB-C22
  • dc.relation.projectID info:eu-repo/grantAgreement/ES/2PE/PID2020-114627RB-I00
  • dc.relation.projectID info:eu-repo/grantAgreement/ES/3PE/PDC2021-121544-I00
  • dc.relation.projectID info:eu-repo/grantAgreement/ES/2PE/PID2020-112821GB-I00
  • dc.rights © 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
  • dc.rights.accessRights info:eu-repo/semantics/openAccess
  • dc.rights.uri http://creativecommons.org/licenses/by/4.0/
  • dc.subject.keyword Antimicrobial peptides
  • dc.subject.keyword Panusin
  • dc.subject.keyword β-defensins
  • dc.title The C-terminus of panusin, a lobster β-defensin, is crucial for optimal antimicrobial activity and serum stability
  • dc.type info:eu-repo/semantics/article
  • dc.type.version info:eu-repo/semantics/publishedVersion

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