An antibacterial novel peptide based on cecropin and MAP-27: Design and characterization

Majed M. Masadeh Anwar E. Abu AL-Kahsi Razan Haddad Mohammad Alsaggar Karem H. Alzoubi Salsabeel H. Sabi Nasr Alrabadi   

Open Access   

Published:  Nov 23, 2022

DOI: 10.7324/JAPS.2023.62382
Abstract

Antibiotic development has passed its peak, yet new antibiotics have never been more necessary in relation to the rising antibiotic resistance rate. The high prevalence of multidrug resistance (MDR) bacterial infections, due to microbes’ ability to overcome antibiotics, is a huge burden to the healthcare system. The aim of this study was to design a novel altered hybrid peptide named HEA-9 from the natural parent peptides BAMP-27 and cecropin A with improved activity and selectivity. HEA-9 was rationally designed by hybridizing the active residues of the parent peptides. This was followed by an amino acid modification to enhance the physicochemical properties of HEA-9, which were evaluated using in silico tools. Thereafter, the in vitro antibacterial activities of HEA-9 against sensitive and MDR strains of Gram-negative and Gram-positive bacteria were measured. Furthermore, the antibiofilm activities against MDR bacteria were evaluated. Moreover, synergistic experiments with four conventional antibiotics were conducted against all tested bacteria. Finally, we used Vero cells to assess HEA-9/associated cytotoxicity incorporated into mammalian cells, and we examined its hemolytic activity on erythrocytes. HEA-9 expressed extensive activity against sensitive and MDR strains of Staphylococcus aureus and Escherichia coli bacteria, having a 12.5 μM minimum inhibitory concentration (MIC)/MBC. HEA-9 was also capable of eradicating biofilms, with reported minimal biofilm eradication concentrations of 100 and 25 μM for MDR E. coli and MDR S. aureus, respectively. Also, HEA-9 demonstrated superior toxicity profiles against erythrocyte cells and Vero cells. Combinations of HEA-9 with conventional antibiotics resulted in a considerable enhancement in the antibacterial activity of the combined drugs. Interestingly, the MIC of HEA-9 in conjunction with traditional antibiotics decreased up to 0.098 μM in certain situations. In conclusion, the HEA-9 peptide has shown improved activity and selectivity either alone or in combination with conventional antibiotics, making it a promising candidate for treating MDR bacterial infections.


Keyword:     Antimicrobial peptides rational design hybridization antimicrobial resistance MDR bacteria antibiofilm activity synergism


Citation:

Masadeh MM, AL-Kahsi AEA, Haddad R, Alsaggar M, Alzoubi KH, Sabi SH, Alrabadi N. An antibacterial novel peptide based on cecropin and MAP-27: Design and characterization. J Appl Pharm Sci, 2022. https://doi.org/10.7324/JAPS.2023.62382

Copyright: © The Author(s). This is an open-access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Reference

Albsoul-Younes A, Wazaify M, Yousef AM, Tahaineh L. Abuse and misuse of prescription and nonprescription drugs sold in community pharmacies in Jordan. Subst Use Misuse, 2010; 45(9):1319-29. https://doi.org/10.3109/10826080802490683

Almaaytah A, Zhou M, Wang L, Chen T, Walker B, Shaw C. Antimicrobial/cytolytic peptides from the venom of the North African scorpion, Androctonus amoreuxi: biochemical and functional characterization of natural peptides and a single site-substituted analog. Peptides, 2012; 35(2):291-9. https://doi.org/10.1016/j.peptides.2012.03.016

Andersson DI, Hughes D, Kubicek-Sutherland JZ. Mechanisms and consequences of bacterial resistance to antimicrobial peptides. Drug Resist Updates, 2016; 26:43-57. https://doi.org/10.1016/j.drup.2016.04.002

Azmi F, Skwarczynski M, Toth I. Towards the development of synthetic antibiotics: designs inspired by natural antimicrobial peptides. Curr Med Chem, 2016; 23(41):4610-24. https://doi.org/10.2174/0929867323666160825162435

Baeumlisberger D, Arrey TN, Rietschel B, Rohmer M, Papasotiriou DG, Mueller B, Beckhaus T, Karas M. Labeling elastase digests with TMT: informational gain by identification of poorly detectable peptides with MALDI-TOF/TOF mass spectrometry. Proteomics, 2010; 10(21):3905-9. https://doi.org/10.1002/pmic.201000288

Bauer K, Struyve M, Bosch D, Benz R, Tommassen J. One single lysine residue is responsible for the special interaction between polyphosphate and the outer membrane porin PhoE of Escherichia coli. Journal of Biological Chemistry. 1989 Oct 5;264(28):16393-8. https://doi.org/10.1016/S0021-9258(19)84719-1

Beaufays J, Lins L, Thomas A, Brasseur R. In silico predictions of 3D structures of linear and cyclic peptides with natural and non-proteinogenic residues. J Pept Sci, 2012; 18(1):17-24. Cama J, Leszczynski R, Tang P, Khalid A, Lok V, Dowson C, Ebata A. To push or to pull? In a post-COVID world, supporting and incentivizing antimicrobial drug development must become a governmental priority. ACS Infect Dis, 2021; 7:2029-42. https://doi.org/10.1021/acsinfecdis.0c00681

Chen Y, Guarnieri MT, Vasil AI, Vasil ML, Mant CT, Hodges RS. Role of peptide hydrophobicity in the mechanism of action of alpha-helical antimicrobial peptides. Antimicrob Agents Chemother, 2007; 51(4):1398-406. https://doi.org/10.1128/AAC.00925-06

Ceri H, Olson M, Morck D, Storey D, Read R, Buret A, Olson B. The MBEC assay system: multiple equivalent biofilms for antibiotic and biocide susceptibility testing. Methods Enzymol, 2001; 337:377-85. https://doi.org/10.1016/S0076-6879(01)37026-X

CLSI. Performance standards for antimicrobial susceptibility testing; twenty-fourth informational supplement. CLSI, Wayne, PA, 2014.

Combet C, Blanchet C, Geourjon C, Deleage G. NPS@: network protein sequence analysis. Trends Biochem Sci, 2000; 25(3):147-50. https://doi.org/10.1016/S0968-0004(99)01540-6

Ciumac D, Gong H, Hu X, Lu JR. Membrane targeting cationic antimicrobial peptides. J Colloid Interface Sci, 2019; 537:163-85. https://doi.org/10.1016/j.jcis.2018.10.103

Donlan RM, Costerton JW. Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev, 2002; 15(2):167-93. https://doi.org/10.1128/CMR.15.2.167-193.2002

Eckert R. Road to clinical efficacy: challenges and novel strategies for antimicrobial peptide development. Future Microbiol, 2011; 6(6):635-51. https://doi.org/10.2217/fmb.11.27

Epand RM, Epand RF. Lipid domains in bacterial membranes and the action of antimicrobial agents. Biochim Biophys Acta, 2009; 1788(1):289-94. https://doi.org/10.1016/j.bbamem.2008.08.023

Gupta UK, Mahanta S, Paul S. In silico design of small peptide-based Hsp90 inhibitor: a novel anticancer agent. Med Hypotheses, 2013; 81(5):853-61. https://doi.org/10.1016/j.mehy.2013.08.006

Fernandes P. Antibacterial discovery and development-the failure of success? Nat Biotechnol, 2006; 24(12):1497-503. https://doi.org/10.1038/nbt1206-1497

Fjell CD, Hiss JA, Hancock RE, Schneider G. Designing antimicrobial peptides: form follows function. Nat Rev Drug Discov, 2012; 11(1):37-51. https://doi.org/10.1038/nrd3591

Gasteiger E, Hoogland C, Gattiker A, Wilkins MR, Appel RD, Bairoch A. Protein identification and analysis tools on the ExPASy server. In: Walker JM (Ed.). The proteomics protocols handbook, Humana Press, Totowa, NJ, pp 571-607, 2005. https://doi.org/10.1385/1-59259-890-0:571

Gautier R, Douguet D, Antonny B, Drin G. HELIQUEST: a web server to screen sequences with specific α-helical properties. Bioinformatics, 2008; 24(18):2101-2. https://doi.org/10.1093/bioinformatics/btn392

Gelband H, Molly Miller P, Pant S, Gandra S, Levinson J, Barter D, White A, Laxminarayan R. The state of the world's antibiotics 2015. Wound Healing South Afr, 2015; 8(2):30-4.

Gennaro R, Zanetti M. Structural features and biological activities of the cathelicidin-derived antimicrobial peptides. Peptide Sci, 2000; 55(1):31-49. https://doi.org/10.1002/1097-0282(2000)55:1<31::AID-BIP40>3.0.CO;2-9

Giangaspero A, Sandri L, Tossi A. Amphipathic α helical antimicrobial peptides. A systematic study of the effects of structural and physical properties on biological activity. Eur J Biochem, 2001; 268(21):5589-600. https://doi.org/10.1046/j.1432-1033.2001.02494.x

Gill EE, Franco OL, Hancock RE. Antibiotic adjuvants: diverse strategies for controlling drug-resistant pathogens. Chem Biol Drug Des, 2015; 85(1):56-78. https://doi.org/10.1111/cbdd.12478

Harding CM, Hennon SW, Feldman MF. Uncovering the mechanisms of Acinetobacter baumannii virulence. Nat Rev Microbiol, 2018; 16(2):91-102. https://doi.org/10.1038/nrmicro.2017.148

Huang Y, Huang J, Chen Y. Alpha-helical cationic antimicrobial peptides: relationships of structure and function. Protein Cell, 2010; 1(2):143-52. https://doi.org/10.1007/s13238-010-0004-3

Halder A, Karmakar S. An evidence of pores in phospholipid membrane induced by an antimicrobial peptide NK-2. Biophy Chem, 2022; 282:106759. https://doi.org/10.1016/j.bpc.2022.106759

Huan Y, Kong Q, Mou H, Yi H. Antimicrobial peptides: classification, design, application and research progress in multiple fields. Front Microbiol, 2020; 11:582779. https://doi.org/10.3389/fmicb.2020.582779

Janocha S, Bichet A, Zöllner A, Bernhardt R. Substitution of lysine with glutamic acid at position 193 in bovine CYP11A1 significantly affects protein oligomerization and solubility but not enzymatic activity. Biochimica et Biophysica Acta (BBA)-Proteins and Proteomics. 2011 Jan 1;1814(1):126-31. https://doi.org/10.1016/j.bbapap.2010.06.002

Jiang Y, Chen Y, Song Z, Tan Z, Cheng J. Recent advances in design of antimicrobial peptides and polypeptides toward clinical translation. Adv Drug Deliv Rev, 2021; 170:261-80. https://doi.org/10.1016/j.addr.2020.12.016

Jureti? D, Sonavane Y, Ili? N, Gajski G, Goi?-Bariši? I, Tonki? M, Kozic M, Maravi? A, Pellay FX, Zorani? L. Designed peptide with a flexible central motif from ranatuerins adapts its conformation to bacterial membranes. Biochim Biophys Acta Biomembr, 2018; 1860(12):2655-68. https://doi.org/10.1016/j.bbamem.2018.10.005

Kardani K, Bolhassani A. Exploring novel and potent cell penetrating peptides in the proteome of SARS-COV-2 using bioinformatics approaches. PLoS One, 2021; 16(2):e0247396. https://doi.org/10.1371/journal.pone.0247396

Klubthawee N, Adisakwattana P, Hanpithakpong W, Somsri S, Aunpad R. A novel, rationally designed, hybrid antimicrobial peptide, inspired by cathelicidin and aurein, exhibits membrane-active mechanisms against Pseudomonas aeruginosa. Sci Rep, 2020; 10(1):1-17. https://doi.org/10.1038/s41598-020-65688-5

Ko SJ, Park E, Asandei A, Choi JY, Lee SC, Seo CH, Luchian T, Park Y. Bee venom-derived antimicrobial peptide melectin has broad-spectrum potency, cell selectivity, and salt-resistant properties. Sci Rep, 2020; 10(1):10145. https://doi.org/10.1038/s41598-020-66995-7

Kourkouta L, Koukourikos K, Iliadis C, Plati P, Dimitriadou A. History of antibiotics. Sumerian J Med Healthcare, 2018; 1:51-5.

Kumar P, Kizhakkedathu JN, Straus SK. Antimicrobial peptides: diversity, mechanism of action and strategies to improve the activity and biocompatibility in vivo. Biomolecules, 2018; 8(1). https://doi.org/10.3390/biom8010004

Lange AB. Crustacean cardioactive peptide in the Chagas' disease vector, Rhodnius prolixus: presence, distribution and physiological effects. Gen Comp Endocrinol, 2011; 174(1):36-43. https://doi.org/10.1016/j.ygcen.2011.08.007

Lee E, Jeong KW, Lee J, Shin A, Kim JK, Lee J, Lee DG, Kim Y. Structure-activity relationships of cecropin-like peptides and their interactions with phospholipid membrane. BMB Rep, 2013; 46(5):282. https://doi.org/10.5483/BMBRep.2013.46.5.252

Lee EK, Kim YC, Nan YH, Shin SY. Cell selectivity, mechanism of action and LPS-neutralizing activity of bovine myeloid antimicrobial peptide-18 (BMAP-18) and its analogs. Peptides, 2011; 32(6):1123-30. https://doi.org/10.1016/j.peptides.2011.03.024

Lewis K. Platforms for antibiotic discovery. Nat Rev Drug Discov, 2013; 12(5):371-87. https://doi.org/10.1038/nrd3975

Liévin-Le Moal V, Servin AL. The front line of enteric host defense against unwelcome intrusion of harmful microorganisms: mucins, antimicrobial peptides, and microbiota. Clin Microbiol Rev, 2006; 19(2):315-37. https://doi.org/10.1128/CMR.19.2.315-337.2006

Lundstedt E, Kahne D, Ruiz N. Assembly and maintenance of lipids at the bacterial outer membrane. Chem Rev, 2021; 121(9):5098-123. https://doi.org/10.1021/acs.chemrev.0c00587

Mahlapuu M, Håkansson J, Ringstad L, Björn C. Antimicrobial peptides: an emerging category of therapeutic agents. Front Cell Infect Microbiol, 2016; 6:194. https://doi.org/10.3389/fcimb.2016.00194

Malanovic N, Lohner K. Antimicrobial peptides targeting Gram-positive bacteria. Pharmaceuticals, 2016; 9(3):59. https://doi.org/10.3390/ph9030059

Masadeh M, Ayyad A, Haddad R, Alsaggar M, Alzoubi K, Alrabadi N. Functional and toxicological evaluation of the MAA-41: a novel rationally designed antimicrobial peptide using hybridization and modification methods from LL-37 and BMAP-28. Curr Pharm Des, 2022; 28:2177-88. https://doi.org/10.2174/1381612828666220705150817

McDermott P, Zhao S, Wagner D, Simjee S, Walker R, White D. The food safety perspective of antibiotic resistance. Anim Biotechnol, 2002; 13(1):71-84. https://doi.org/10.1081/ABIO-120005771

Mohr KI. History of antibiotics research. How to overcome the antibiotic crisis. Curr Top Microbiol Immunol, 2016; 398:237-72. https://doi.org/10.1007/82_2016_499

Moore AJ, Beazley WD, Bibby MC, Devine DA. Antimicrobial activity of cecropins. J Antimicrob Chemother, 1996; 37(6):1077-89. https://doi.org/10.1093/jac/37.6.1077

Moravej H, Moravej Z, Yazdanparast M, Heiat M, Mirhosseini A, Moosazadeh Moghaddam M, Mirnejad R. Antimicrobial peptides: features, action, and their resistance mechanisms in bacteria. Microb Drug Resist, 2018; 24(6):747-67. https://doi.org/10.1089/mdr.2017.0392

Neu HC, Gootz TD. Antimicrobial chemotherapy. Medical microbiology. 4th edition, University of Texas Medical Branch, Galveston, TX, 1996.

Ron?evi? T, Puizina J, Tossi A. Antimicrobial peptides as anti-infective agents in pre-post-antibiotic era? Int J Mol Sci, 2019; 20(22). https://doi.org/10.3390/ijms20225713

Roy S, Maheshwari N, Chauhan R, Sen NK, Sharma A. Structure prediction and functional characterization of secondary metabolite proteins of Ocimum. Bioinformation, 2011; 6(8):315-9. https://doi.org/10.6026/97320630006315

Shields RK, Chen L, Cheng S, Chavda KD, Press EG, Snyder A, Pandey R, Doi Y, Kreiswirth BN, Nguyen M, Clancy CJ. Emergence of ceftazidime-avibactam resistance due to plasmid-borne blaKPC-3 mutations during treatment of carbapenem-resistant Klebsiella pneumoniae infections. Antimicrob Agents Chemother, 2017; 61(3):e02097-16. https://doi.org/10.1128/AAC.02097-16

Simões M, Simões LC, Vieira MJ. A review of current and emergent biofilm control strategies. LWT Food Sci Technol, 2010; 43(4):573-83. https://doi.org/10.1016/j.lwt.2009.12.008

Skerlavaj B, Gennaro R, Bagella L, Merluzzi L, Risso A, Zanetti M. Biological characterization of two novel cathelicidin-derived peptides and identification of structural requirements for their antimicrobial and cell lytic activities. J Biol Chem, 1996; 271(45):28375-81. https://doi.org/10.1074/jbc.271.45.28375

Toke O. Antimicrobial peptides: new candidates in the fight against bacterial infections. Peptide Sci, 2005; 80(6):717-35. https://doi.org/10.1002/bip.20286

Tong SYC, Davis JS, Eichenberger E, Holland TL, Fowler VG. Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management. Clin Microbiol Rev, 2015; 28:603-61. https://doi.org/10.1128/CMR.00134-14

Ventola CL. The antibiotic resistance crisis: part 1: causes and threats. Pharm Ther, 2015; 40(4):277.

Walsh C. Antibiotics: actions, origins, resistance. American Society for Microbiology (ASM), Washington, DC, 2003. https://doi.org/10.1128/9781555817886

Wang G, Li X, Wang Z. APD3: the antimicrobial peptide database as a tool for research and education. Nucleic Acids Res, 2016a; 44(D1):D1087-93. https://doi.org/10.1093/nar/gkv1278

Wang S, Zeng X, Yang Q, Qiao S. Antimicrobial peptides as potential alternatives to antibiotics in food animal industry. Int J Mol Sci, 2016b; 17(5):603. https://doi.org/10.3390/ijms17050603

Wei XB, Wu RJ, Si DY, Liao XD, Zhang LL, Zhang RJ. Novel hybrid peptide cecropin A (1-8)-LL37 (17-30) with potential antibacterial activity. Int J Mol Sci, 2016; 17(7):983. https://doi.org/10.3390/ijms17070983

Wu R, Wang Q, Zheng Z, Zhao L, Shang Y, Wei X, Liao X, Zhang R. Design, characterization and expression of a novel hybrid peptides melittin (1-13)-LL37 (17-30). Mol Biol Rep, 2014; 41(7):4163-9. https://doi.org/10.1007/s11033-013-2900-0

Yang S, Lee CW, Kim HJ, Jung HH, Kim JI, Shin SY, Shin SH. Structural analysis and mode of action of BMAP-27, a cathelicidin-derived antimicrobial peptide. Peptides, 2019; 118:170106. https://doi.org/10.1016/j.peptides.2019.170106

Zhang Y, Liu Y, Sun Y, Liu Q, Wang X, Li Z, Hao J. In vitro synergistic activities of antimicrobial peptide brevinin-2CE with five kinds of antibiotics against multidrug-resistant clinical isolates. Curr Microbiol, 2014; 68(6):685-92. https://doi.org/10.1007/s00284-014-0529-4

Zhang Y. I-TASSER server for protein 3D structure prediction. BMC Bioinformatics, 2008; 9(1):40. https://doi.org/10.1186/1471-2105-9-40

Zharkova MS, Orlov DS, Golubeva OY, Chakchir OB, Eliseev IE, Grinchuk TM, Shamova OV. Application of antimicrobial peptides of the innate immune system in combination with conventional antibiotics-a novel way to combat antibiotic resistance? Front Cell Infect Microbiol, 2019; 9:128. https://doi.org/10.3389/fcimb.2019.00128

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