Abstract
Antimicrobial peptides (AMPs) are a crucial component of the natural defense system that the host employs to protect itself against invading pathogens. PMAP-23, a cathelicidin-derived AMP, has potent and broad-spectrum antimicrobial activity. Our earlier studies led us to hypothesize that PMAP-23 adopts a dynamic helix–hinge–helix structure, initially attaching to membrane surfaces through the N-helix and subsequently inserting the C-helix into the lipid bilayer. Here, we rationally designed PMAP-NC with increased amphipathicity and hydrophobicity in the N- and C-helix, respectively, based on the hypothesis of the interaction of PMAP-23 with membranes. Compared to the parental PMAP-23, PMAP-NC showed two–eightfold improved bactericidal activity against both Gram-positive and Gram-negative strains with fast killing kinetics. Fluorescence studies demonstrated that PMAP-NC largely disrupted membrane integrity, indicating that efficiency and kinetics of bacterial killing are associated with the membrane permeabilization. Interestingly, PMAP-NC exhibited much better anticancer activity against tumor cells than PMAP-23 but displayed low hemolytic activity against human erythrocytes. Collectively, our findings suggest that PMAP-NC, with the structural arrangement of an amphipathic helix–hinge–hydrophobic helix that plays a critical role in rapid and efficient membrane permeabilization, can be an attractive candidate for novel antimicrobial and/or anticancer drugs.
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References
Bevers EM, Williamson PL (2016) Getting to the outer leaflet: physiology of phosphatidylserine exposure at the plasma membrane. Physiol Rev 96(2):605–645. https://doi.org/10.1152/physrev.00020.2015
Bhattacharjya S (2016) NMR structures and interactions of antimicrobial peptides with lipopolysaccharide: connecting structures to functions. Curr Top Med Chem 16(1):4–15. https://doi.org/10.2174/1568026615666150703121943
Bhattacharjya S, Mohid SA, Bhunia A (2022) Atomic-resolution structures and mode of action of clinically relevant antimicrobial peptides. Int J Mol Sci. https://doi.org/10.3390/ijms23094558
Bhunia A, Ramamoorthy A, Bhattacharjya S (2009) Helical hairpin structure of a potent antimicrobial peptide MSI-594 in lipopolysaccharide micelles by NMR spectroscopy. Chemistry 15(9):2036–2040. https://doi.org/10.1002/chem.200802635
Bhunia A, Domadia PN, Torres J, Hallock KJ, Ramamoorthy A, Bhattacharjya S (2010) NMR structure of pardaxin, a pore-forming antimicrobial peptide, in lipopolysaccharide micelles: mechanism of outer membrane permeabilization. J Biol Chem 285(6):3883–3895. https://doi.org/10.1074/jbc.M109.065672
Biswaro LS, da Costa Sousa MG, Rezende TMB, Dias SC, Franco OL (2018) Antimicrobial peptides and nanotechnology, recent advances and challenges. Front Microbiol 9:855. https://doi.org/10.3389/fmicb.2018.00855
Browne K, Chakraborty S, Chen RX, Willcox MDP, Black DS, Walsh WR, Kumar N (2020) A new era of antibiotics: the clinical potential of antimicrobial peptides. Int J Mol Sci 21(19):7047. https://doi.org/10.3390/Ijms21197047
de Breij A, Riool M, Cordfunke RA, Malanovic N, de Boer L, Koning RI, Ravensbergen E, Franken M, van der Heijde T, Boekema BK, Kwakman PHS, Kamp N, El Ghalbzouri A, Lohner K, Zaat SAJ, Drijfhout JW, Nibbering PH (2018) The antimicrobial peptide SAAP-148 combats drug-resistant bacteria and biofilms. Sci Transl Med. https://doi.org/10.1126/scitranslmed.aan4044
Deslouches B, Di YP (2017) Antimicrobial peptides with selective antitumor mechanisms: prospect for anticancer applications. Oncotarget 8(28):46635–46651. https://doi.org/10.18632/oncotarget.16743
Domadia PN, Bhunia A, Ramamoorthy A, Bhattacharjya S (2010) Structure, interactions, and antibacterial activities of MSI-594 derived mutant peptide MSI-594F5A in lipopolysaccharide micelles: role of the helical hairpin conformation in outer-membrane permeabilization. J Am Chem Soc 132(51):18417–18428. https://doi.org/10.1021/ja1083255
Fjell CD, Hiss JA, Hancock RE, Schneider G (2011) Designing antimicrobial peptides: form follows function. Nat Rev Drug Discov 11(1):37–51. https://doi.org/10.1038/nrd3591[pii]
Gaspar D, Veiga AS, Castanho MA (2013) From antimicrobial to anticancer peptides. A Review Front Microbiol 4:294. https://doi.org/10.3389/fmicb.2013.00294
Guha S, Ghimire J, Wu E, Wimley WC (2019) Mechanistic landscape of membrane-permeabilizing peptides. Chem Rev. https://doi.org/10.1021/acs.chemrev.8b00520
Haney EF, Straus SK, Hancock REW (2019) Reassessing the host defense peptide landscape. Front Chem 7:43. https://doi.org/10.3389/fchem.2019.00043
Hilchie AL, Wuerth K, Hancock RE (2013) Immune modulation by multifaceted cationic host defense (antimicrobial) peptides. Nat Chem Biol 9(12):761–768. https://doi.org/10.1038/nchembio.1393
Jafari A, Babajani A, Sarrami Forooshani R, Yazdani M, Rezaei-Tavirani M (2022) Clinical applications and anticancer effects of antimicrobial peptides: from bench to bedside. Front Oncol 12:819563. https://doi.org/10.3389/fonc.2022.819563
Kobayashi S, Takeshima K, Park CB, Kim SC, Matsuzaki K (2000) Interactions of the novel antimicrobial peptide buforin 2 with lipid bilayers: proline as a translocation promoting factor. Biochemistry Us 39(29):8648–8654 (bi0004549[pii])
Kosciuczuk EM, Lisowski P, Jarczak J, Strzalkowska N, Jozwik A, Horbanczuk J, Krzyzewski J, Zwierzchowski L, Bagnicka E (2012) Cathelicidins: family of antimicrobial peptides. a review. Mol Biol Rep 39(12):10957–10970. https://doi.org/10.1007/s11033-012-1997-x
Kumar P, Kizhakkedathu JN, Straus SK (2018) Antimicrobial peptides: diversity mechanism of action and strategies to improve the activity and biocompatibility in vivo. Biomolecules. https://doi.org/10.3390/biom8010004
Lee TH, Hall KN, Aguilar MI (2016) Antimicrobial peptide structure and mechanism of action: a focus on the role of membrane structure. Curr Top Med Chem 16(1):25–39
Lee H, Lim SI, Shin SH, Lim Y, Koh JW, Yang S (2019) Conjugation of cell-penetrating peptides to antimicrobial peptides enhances antibacterial activity. ACS Omega 4(13):15694–15701. https://doi.org/10.1021/acsomega.9b02278
Lee H, Yang S, Shin SY (2020) Improved cell selectivity of symmetric alpha-helical peptides derived from Trp-Rich antimicrobial peptides. B Korean Chem Soc 41(9):930–936. https://doi.org/10.1002/bkcs.12091
Liu Y, Shen T, Chen L, Zhou J, Wang C (2020) Analogs of the cathelicidin-derived antimicrobial peptide PMAP-23 exhibit improved stability and antibacterial activity. Probiotics Antimicrob Proteins. https://doi.org/10.1007/s12602-020-09686-z
Liu Y, Shen T, Chen L, Zhou J, Wang C (2021) Analogs of the cathelicidin-derived antimicrobial peptide PMAP-23 exhibit improved stability and antibacterial activity. Probiotics Antimicrob Proteins 13(1):273–286. https://doi.org/10.1007/s12602-020-09686-z
Lohner K (2016) Membrane-active antimicrobial peptides as template structures for novel antibiotic agents. Curr Top Med Chem 17:508
Mahlapuu M, Hakansson J, Ringstad L, Bjorn C (2016) Antimicrobial peptides: an emerging category of therapeutic agents. Front Cell Infect Microbiol 6:194. https://doi.org/10.3389/fcimb.2016.00194
Matsuzaki K (2019) Membrane Permeabilization Mechanisms. Adv Exp Med Biol 1117:9–16. https://doi.org/10.1007/978-981-13-3588-4_2
Park K, Oh D, Shin SY, Hahm KS, Kim Y (2002) Structural studies of porcine myeloid antibacterial peptide PMAP-23 and its analogues in DPC micelles by NMR spectroscopy. Biochem Biophys Res Commun 290(1):204–212. https://doi.org/10.1006/bbrc.2001.6173
Pfalzgraff A, Brandenburg K, Weindl G (2018) Antimicrobial peptides and their therapeutic potential for bacterial skin infections and wounds. Front Pharmacol 9:281. https://doi.org/10.3389/Fphar.2018.00281
Sani MA, Separovic F (2016) How membrane-active peptides get into lipid membranes. Acc Chem Res 49(6):1130–1138. https://doi.org/10.1021/acs.accounts.6b00074
Scott RW, DeGrado WF, Tew GN (2008) De novo designed synthetic mimics of antimicrobial peptides. Curr Opin Biotechnol 19(6):620–627. https://doi.org/10.1016/j.copbio.2008.10.013S0958-1669(08)00143-2[pii]
Shin HJ, Yang S, Lim Y (2021) Antibiotic susceptibility of Staphylococcus aureus with different degrees of biofilm formation. J Anal Sci Technol 12(1):41. https://doi.org/10.1186/S40543-021-00294-2
Stone TA, Cole GB, Ravamehr-Lake D, Nguyen HQ, Khan F, Sharpe S, Deber CM (2019) Positive charge patterning and hydrophobicity of membrane-active antimicrobial peptides as determinants of activity, toxicity, and pharmacokinetic stability. J Med Chem. https://doi.org/10.1021/acs.jmedchem.9b00657
Tossi A (2011) Design and engineering strategies for synthetic antimicrobial peptides. Prokaryotic Antimicrob Pept. https://doi.org/10.1007/978-1-4419-7692-5_6
Veldhuizen EJA, Scheenstra MR, Tjeerdsma-van Bokhoven JLM, Coorens M, Schneider VAF, Bikker FJ, van Dijk A, Haagsman HP (2017) Antimicrobial and immunomodulatory activity of PMAP-23 derived peptides. Protein Pept Lett 24(7):609–616. https://doi.org/10.2174/0929866524666170428150925
Wimley WC (2010) Describing the mechanism of antimicrobial peptide action with the interfacial activity model. ACS Chem Biol 5(10):905–917. https://doi.org/10.1021/cb1001558
Yang ST, Jeon JH, Kim Y, Shin SY, Hahm KS, Kim JI (2006a) Possible role of a PXXP central hinge in the antibacterial activity and membrane interaction of PMAP-23, a member of cathelicidin family. Biochemistry 45(6):1775–1784. https://doi.org/10.1021/bi051524k
Yang ST, Lee JY, Kim HJ, Eu YJ, Shin SY, Hahm KS, Kim JI (2006b) Contribution of a central proline in model amphipathic alpha-helical peptides to self-association, interaction with phospholipids, and antimicrobial mode of action. FEBS J 273(17):4040–4054. https://doi.org/10.1111/j.1742-4658.2006.05407.x. (EJB5407 [pii])
Yang ST, Shin SY, Hahm KS, Kim JI (2006c) Design of perfectly symmetric Trp-rich peptides with potent and broad-spectrum antimicrobial activities. Int J Antimicrob Agents 27(4):325–330. https://doi.org/10.1016/j.ijantimicag.2005.11.014. (S0924-8579(06)00027-6 [pii])
Yang ST, Shin SY, Hahm KS, Kim JI (2006d) Different modes in antibiotic action of tritrpticin analogs, cathelicidin-derived Trp-rich and Pro/Arg-rich peptides. Biochim Biophys Acta 1758(10):1580–1586. https://doi.org/10.1016/j.bbamem.2006.06.007. (S0005-2736(06)00217-3 [pii])
Yang ST, Kim JI, Shin SY (2009) Effect of dimerization of a beta-turn antimicrobial peptide, PST13-RK, on antimicrobial activity and mammalian cell toxicity. Biotechnol Lett 31(2):233–237. https://doi.org/10.1007/s10529-008-9848-5
Yang S, Lee CW, Kim HJ, Jung HH, Kim JI, Shin SY, Shin SH (2019) Structural analysis and mode of action of BMAP-27, a cathelicidin-derived antimicrobial peptide. Peptides 118:170106. https://doi.org/10.1016/j.peptides.2019.170106
Yang ST, Shin SY, Shin SH (2021) The central PXXP motif is crucial for PMAP-23 translocation across the lipid bilayer. Int J Mol Sci 22(18):9752. https://doi.org/10.3390/Ijms22189752
Zanetti M (2005) The role of cathelicidins in the innate host defenses of mammals. Curr Issues Mol Biol 7(2):179–196
Zhang LJ, Gallo RL (2016) Antimicrobial peptides. Curr Biol 26(1):R14-19. https://doi.org/10.1016/j.cub.2015.11.017
Zhang L, Dhillon P, Yan H, Farmer S, Hancock RE (2000) Interactions of bacterial cationic peptide antibiotics with outer and cytoplasmic membranes of pseudomonas aeruginosa. Antimicrob Agents Chemother 44(12):3317–3321
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This study was supported by a research fund from Chosun University (2021).
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S.S. and S.Y. designed the experiments, and H.L. conducted the experimental work. All the authors collectively assessed the results and wrote the manuscript.
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Lee, H., Shin, SH. & Yang, S. Rationally designed PMAP-23 derivatives with enhanced bactericidal and anticancer activity based on the molecular mechanism of peptide–membrane interactions. Amino Acids 55, 1013–1022 (2023). https://doi.org/10.1007/s00726-023-03290-5
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DOI: https://doi.org/10.1007/s00726-023-03290-5