A novel antimicrobial peptide found in Pelophylax nigromaculatus
Tóm tắt
Many active peptides have been found in frog skin secretions. In this paper, our research focused on Pelophylax nigromaculatus and found a broad-spectrum antimicrobial peptide Nigrocin-PN based on the molecular cloning technique. Thereafter, the “Rana box” function was briefly studied by two mutated peptides (Nigrocin-M1 and Nigrocin-M2). Furthermore, in vitro and in vivo assays were used to characterize the peptide’s biofunctions, and the peptide’s function in treating multidrug-resistant pathogens was also studied. Nigrocin-PN not only displayed potent antimicrobial abilities in vitro but also significantly ameliorated pulmonary inflammation induced by Klebsiella pneumoniae in vivo. By comparing, leucine-substituted analogue Nigrocin-M1 only displayed bactericidal abilities towards gram-positive bacteria, while the shorter analogue Nigrocin-M2 lost this function. More strikingly, Nigrocin-PN exhibited synergistic effects with commonly used antibiotics; in vitro evolution experiments revealed that coadministration between Nigrocin-PN and ampicillin could delay Staphylococcus aureus antibiotic resistance acquisition. Kinetics and morphology studies indicate that antibacterial mechanisms involved membrane destruction. Furthermore, toxicities and anticancer abilities of these peptides were also studied; compared to two analogues, Nigrocin-PN showed mild haemolytic activity and indistinctive cytotoxicity towards normal cell lines HMEC-1 and HaCaT. A broad-spectrum antimicrobial peptide Nigrocin-PN was discovered from the skin secretion of Pelophylax nigromaculatus. Structurally, “Rana box” played a crucial role in reducing toxicities without compromising antibacterial abilities, and Nigrocin-PN could be a desired therapeutic candidate.
• For AMPs, disulphide bond can affect their biofunction and cytotoxicity. • Frog skin secretion is a reservoir to delve valuable peptides. • AMPs-antibiotics coadministration could be a strategy to delay drug resistance.
Tài liệu tham khảo
Pandit G, Biswas K, Ghosh S, Debnath S, Bidkar AP, Satpati P, Bhunia A, Chatterjee S (2020) Rationally designed antimicrobial peptides: insight into the mechanism of eleven residue peptides against microbial infections. Biochim Biophys Acta Biomembr 1862(4):183177
Brown ED, Wright GD (2016) Antibacterial drug discovery in the resistance era. Nature 529(7586):336–343
Dickey SW, Cheung GYC, Otto M (2017) Different drugs for bad bugs: antivirulence strategies in the age of antibiotic resistance. Nat Rev Drug Discov 16(7):457–471
Li W, Separovic F, O’Brien-Simpson NM, Wade JD (2021) Chemically modified and conjugated antimicrobial peptides against superbugs. Chem Soc Rev 50(8):4932–4973
Dever LA, Dermody TS (1991) Mechanisms of bacterial resistance to antibiotics. Arch Intern Med 151(5):886–895
Kumar P, Kizhakkedathu JN, Straus SK (2018) Antimicrobial peptides: diversity, mechanism of action and strategies to improve the activity and biocompatibility in vivo. Biomolecules 8(1):24
Guilhelmelli F, Vilela N, Albuquerque P, da Derengowski LS, Silva-Pereira I, Kyaw CM (2013) Antibiotic development challenges: the various mechanisms of action of antimicrobial peptides and of bacterial resistance. Front Microbiol 4:353
Brogden KA (2005) Antimicrobial peptides: pore formers or metabolic inhibitors in bacteria? Nat Rev Microbiol 3(3):238–250
Afacan NJ, Yeung AT, Pena OM, Hancock RE (2012) Therapeutic potential of host defense peptides in antibiotic-resistant infections. Curr Pharm Des 18(6):807–819
Yonezawa A, Kuwahara J, Fujii N, Sugiura Y (1992) Binding of tachyplesin I to DNA revealed by footprinting analysis: significant contribution of secondary structure to DNA binding and implication for biological action. Biochemistry 31(11):2998–3004
Nicolas P, El Amri C (2009) The dermaseptin superfamily: a gene-based combinatorial library of antimicrobial peptides. BBA-Biomembranes 1788(8):1537–1550
Hancock REW, Alford MA, Haney EF (2021) Antibiofilm activity of host defence peptides: complexity provides opportunities. Nat Rev Microbiol 19(12):786–797
Dennison SR, Whittaker M, Harris F, Phoenix DA (2006) Anticancer alpha-helical peptides and structure/function relationships underpinning their interactions with tumour cell membranes. Curr Protein Pept Sci 7(6):487–499
Bao K, Yuan W, Ma C, Yu X, Wang L, Hong M, Xi X, Zhou M, Chen T (2018) Modification targeting the “Rana box” motif of a novel nigrocin peptide from Hylarana latouchii enhances and broadens its potency against multiple bacteria. Front Microbiol 9:2846
Kwon MY, Hong SY, Lee KH (1998) Structure-activity analysis of brevinin 1E amide, an antimicrobial peptide from Rana esculenta. Biochim Biophys Acta 1387(1–2):239–248
Gao Y, Wu D, Wang L, Lin C, Ma C, Xi X, Zhou M, Duan J, Bininda-Emonds ORP, Chen T, Shaw C (2017) Targeted modification of a novel amphibian antimicrobial peptide from Phyllomedusa tarsius to enhance its activity against MRSA and microbial biofilm. Front Microbiol 8:628
Wu D, Gao Y, Tan Y, Liu Y, Wang L, Zhou M, Xi X, Ma C, Bininda-Emonds ORP, Chen T, Shaw C (2018) Discovery of distinctin-like-peptide-PH (DLP-PH) from the skin secretion of Phyllomedusa hypochondrialis, a prototype of a novel family of antimicrobial peptide. Front Microbiol 9:541
Yuan Y, Zai Y, Xi X, Ma C, Wang L, Zhou M, Shaw C, Chen T (2019) A novel membrane-disruptive antimicrobial peptide from frog skin secretion against cystic fibrosis isolates and evaluation of anti-MRSA effect using Galleria mellonella model. Biochim Biophys Acta Gen Subj 1863(5):849–856
Whitmore L, Wallace BA (2008) Protein secondary structure analyses from circular dichroism spectroscopy: methods and reference databases. Biopolymers 89(5):392–400
Whitmore L, Wallace BA (2004) DICHROWEB, an online server for protein secondary structure analyses from circular dichroism spectroscopic data. Nucleic Acids Res 32(Web Server issue):W668-73
Mouton JW, Brown DF, Apfalter P, Canton R, Giske CG, Ivanova M, MacGowan AP, Rodloff A, Soussy CJ, Steinbakk M, Kahlmeter G (2012) The role of pharmacokinetics/pharmacodynamics in setting clinical MIC breakpoints: the EUCAST approach. Clin Microbiol Infect 18(3):E37-45
Segev-Zarko L, Saar-Dover R, Brumfeld V, Mangoni ML, Shai Y (2015) Mechanisms of biofilm inhibition and degradation by antimicrobial peptides. Biochem J 468(2):259–270
Peeters E, Nelis HJ, Coenye T (2008) Comparison of multiple methods for quantification of microbial biofilms grown in microtiter plates. J Microbiol Methods 72(2):157–165
Li T, Wang P, Guo W, Huang X, Tian X, Wu G, Xu B, Li F, Yan C, Liang XJ, Lei H (2019) Natural berberine-based chinese herb medicine assembled nanostructures with modified antibacterial application. ACS Nano 13(6):6770–6781
Huang L, Chen D, Wang L, Lin C, Ma C, Xi X, Chen T, Shaw C, Zhou M (2017) Dermaseptin-PH: a novel peptide with antimicrobial and anticancer activities from the skin secretion of the South American orange-legged leaf frog, Pithecopus (Phyllomedusa) hypochondrialis. Molecules 22(10):1805
Chen X, Zhang L, Wu Y, Wang L, Ma C, Xi X, Bininda-Emonds ORP, Shaw C, Chen T, Zhou M (2018) Evaluation of the bioactivity of a mastoparan peptide from wasp venom and of its analogues designed through targeted engineering. Int J Biol Sci 14(6):599–607
Rishi P, Vij S, Maurya IK, Kaur UJ, Bharati S, Tewari R (2018) Peptides as adjuvants for ampicillin and oxacillin against methicillin-resistant Staphylococcus aureus (MRSA). Microb Pathog 124:11–20
Hsieh MH, Yu CM, Yu VL, Chow JW (1993) Synergy assessed by checkerboard. A critical analysis. Diagn Microbiol Infect Dis 16(4):343–349
Casciaro B, Loffredo MR, Luca V, Verrusio W, Cacciafesta M, Mangoni ML (2018) Esculentin-1a derived antipseudomonal peptides: limited induction of resistance and synergy with Aztreonam. Protein Pept Lett 25(12):1155–1162
Odds FC (2003) Synergy, antagonism, and what the chequerboard puts between them. J Antimicrob Chemother 52(1):1
Suzuki S, Horinouchi T, Furusawa C (2014) Prediction of antibiotic resistance by gene expression profiles. Nat Commun 5:5792
Lazar V, Martins A, Spohn R, Daruka L, Grezal G, Fekete G, Szamel M, Jangir PK, Kintses B, Csorgo B, Nyerges A, Gyorkei A, Kincses A, Der A, Walter FR, Deli MA, Urban E, Hegedus Z, Olajos G, Mehi O, Balint B, Nagy I, Martinek TA, Papp B, Pal C (2018) Antibiotic-resistant bacteria show widespread collateral sensitivity to antimicrobial peptides. Nat Microbiol 3(6):718–731
Ma B, Niu C, Zhou Y, Xue X, Meng J, Luo X, Hou Z (2016) The disulfide bond of the peptide thanatin is dispensible for its antimicrobial activity in vivo and in vitro. Antimicrob Agents Chemother 60(7):4283–4289
Cebrian R, Rodriguez-Cabezas ME, Martin-Escolano R, Rubino S, Garrido-Barros M, Montalban-Lopez M, Rosales MJ, Sanchez-Moreno M, Valdivia E, Martinez-Bueno M, Marin C, Galvez J, Maqueda M (2019) Preclinical studies of toxicity and safety of the AS-48 bacteriocin. J Adv Res 20:129–139
Chen X, Zhang L, Ma C, Zhang Y, Xi X, Wang L, Zhou M, Burrows JF, Chen T (2018) A novel antimicrobial peptide, ranatuerin-2PLx, showing therapeutic potential in inhibiting proliferation of cancer cells. Biosci Rep 38(6):BSR20180710
Fehlbaum P, Bulet P, Chernysh S, Briand JP, Roussel JP, Letellier L, Hetru C, Hoffmann JA (1996) Structure-activity analysis of thanatin, a 21-residue inducible insect defense peptide with sequence homology to frog skin antimicrobial peptides. Proc Natl Acad Sci U S A 93(3):1221–1225
Dash R, Bhattacharjya S (2021) Thanatin: an emerging host defense antimicrobial peptide with multiple modes of action. Int J Mol Sci 22(4):1522
Mohanram H, Bhattacharjya S (2014) Resurrecting inactive antimicrobial peptides from the lipopolysaccharide trap. Antimicrob Agents Chemother 58(4):1987–1996
Bhunia A, Saravanan R, Mohanram H, Mangoni ML, Bhattacharjya S (2011) NMR structures and interactions of temporin-1Tl and temporin-1Tb with lipopolysaccharide micelles: mechanistic insights into outer membrane permeabilization and synergistic activity. J Biol Chem 286(27):24394–24406