Abstract
Tuberculosis has become a cause of worldwide concern; emergence of resistance in various mycobacterial strains has led to an urgent demand for new therapeutic molecules. Antimicrobial peptides have emerged as potential candidates in antimicrobial drug development. We have tested a lead antibacterial peptide LP-23 and its cost-effective mimetic DP-23 for their antimicrobial effects in a non-pathogenic model of Mycobacterium. Their minimum inhibitory concentration (MIC) values against M. smegmatis were calculated by resazurin reduction assay. MIC of both peptidomimetics against M. smegmatis was found to be 6.25 μg/mL. In addition, hemolytic toxicity study of peptidomimetics suggested that the synthesized compounds were selective against bacteria. To better understand stability and mechanism of action of peptidomimetics, serum stability study and SEM analysis were carried out. Peptoid DP-23 was found to be more stable in serum then peptide LP-23, while SEM analysis indicated that these compounds target the cell membrane, impairing the membrane integrity of M. smegmatis. The activity and properties of LP-23 and DP-23 may make them new potential antibacterial agents against tuberculosis.
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References
Arya S, Sethi D, Singh S, Hade MD, Singh V, Raju P, Chodisetti SB, Verma D, Varshney GC, Agrewala JN, Dikshit KL (2013) Truncated hemoglobin, HbN, is post-translationally modified in Mycobacterium tuberculosis and modulates host-pathogen interactions during intracellular infection. J Biol Chem 288(41):29987–29999. https://doi.org/10.1074/jbc.M113.507301
Bolt HL, Eggimann GA, Jahoda CAB, Zuckermann RN, Sharples GJ, Cobb SL (2017) Exploring the links between peptoid antibacterial activity and toxicity. Med Chem Commun 8(5):886–896. https://doi.org/10.1039/C6MD00648E
Bottger R, Hoffmann R, Knappe D (2017) Differential stability of therapeutic peptides with different proteolytic cleavage sites in blood, plasma and serum. PLoS ONE 12(6):e0178943. https://doi.org/10.1371/journal.pone.0178943
Chatrath S, Gupta VK, Dixit A, Garg LC (2016) PE_PGRS30 of Mycobacterium tuberculosis mediates suppression of proinflammatory immune response in macrophages through its PGRS and PE domains. Microbes Infect 18(9):536–542. https://doi.org/10.1016/j.micinf.2016.04.004
Cole AM, Darouiche RO, Legarda D, Connell N, Diamond G (2000) Characterization of a fish antimicrobial peptide: gene expression, subcellular localization, and spectrum of activity. Antimicrob Agents Chemother 44(8):2039–2045. https://doi.org/10.1128/AAC.44.8.2039-2045.2000
Culf AS, Ouellette RJ (2010) Solid-phase synthesis of N-substituted glycine oligomers (alpha-peptoids) and derivatives. Molecules 15(8):5282–5335. https://doi.org/10.3390/molecules15085282
de Menezes YA, Felix-Silva J, da Silva-Junior AA, Rebecchi IM, de Oliveira AS, Uchoa AF, Fernandes-Pedrosa MD (2014) Protein-rich fraction of Cnidoscolus urens (L.) Arthur leaves: enzymatic characterization and procoagulant and fibrinogenolytic activities. Molecules 19(3):3552–3569. https://doi.org/10.3390/molecules19033552
Deng G, Zhang F, Yang S, Kang J, Sha S, Ma Y (2016) Mycobacterium tuberculosis Rv0431 expressed in Mycobacterium smegmatis, a potentially mannosylated protein, mediated the immune evasion of RAW 264.7 macrophages. Microb Pathog 100:285–292. https://doi.org/10.1016/j.micpath.2016.10.013
Gough ME, Graviss EA, May EE (2017) The dynamic immunomodulatory effects of vitamin D3 during Mycobacterium infection. J Innate Immun 23(6):506–523. https://doi.org/10.1177/1753425917719143
Gupta K, Singh S, van Hoek ML (2015) Short, synthetic cationic peptides have antibacterial activity against Mycobacterium smegmatis by forming pores in membrane and synergizing with antibiotics. Antibiotics 4(3):358–378. https://doi.org/10.3390/antibiotics4030358
Hartmann M, Berditsch M, Hawecker J, Ardakani MF, Gerthsen D, Ulrich AS (2010) Damage of the bacterial cell envelope by antimicrobial peptides gramicidin S and PGLa as revealed by transmission and scanning electron microscopy. Antimicrob Agents Chemother 54(8):3132–3142. https://doi.org/10.1128/AAC.00124-10
He Z, De Buck J (2010) Cell wall proteome analysis of Mycobacterium smegmatis strain MC2 155. BMC Microbiol 10(1):121. https://doi.org/10.1186/1471-2180-10-121
Hebert ML, Shah DS, Blake P, Turner JP, Servoss SL (2013) Tunable peptoid microspheres: effects of side chain chemistry and sequence. Org Biomol Chem 11(27):4459–4464. https://doi.org/10.1039/c3ob40561c
Jena P, Mishra B, Leippe M, Hasilik A, Griffiths G, Sonawane A (2011) Membrane-active antimicrobial peptides and human placental lysosomal extracts are highly active against mycobacteria. Peptides 32(5):881–887. https://doi.org/10.1016/j.peptides.2011.03.002
Jenssen H, Aspmo SI (2008) Serum stability of peptides. Methods Mol Biol 494:177–186. https://doi.org/10.1007/978-1-59745-419-3_10
Khara JS, Wang Y, Ke XY, Liu S, Newton SM, Langford PR, Yang YY, Ee PL (2014) Anti-mycobacterial activities of synthetic cationic alpha-helical peptides and their synergism with rifampicin. Biomaterials 35(6):2032–2038. https://doi.org/10.1016/j.biomaterials.2013.11.035
Lohan S, Cameotra SS, Bisht GS (2013) Systematic study of non-natural short cationic lipopeptides as novel broad-spectrum antimicrobial agents. Chem Biol Drug Des 82(5):557–566. https://doi.org/10.1111/cbdd.12182
Lyu Y, Yang Y, Lyu X, Dong N, Shan A (2016) Antimicrobial activity, improved cell selectivity and mode of action of short PMAP-36-derived peptides against bacteria and Candida. Sci Rep 6:27258. https://doi.org/10.1038/srep27258
Merrifield RB (1986) Solid phase synthesis. Science 232:341–347. https://doi.org/10.1126/science.3961484
Mojsoska B, Zuckermann RN, Jenssen H (2015) Structure-activity relationship study of novel peptoids that mimic the structure of antimicrobial peptides. Antimicrob Agents Chemother 59(7):4112–4120. https://doi.org/10.1128/AAC.00237-15
Molchanova N, Hansen PR, Franzyk H (2017) Advances in development of antimicrobial peptidomimetics as potential drugs. Molecules 22(9):1430. https://doi.org/10.3390/molecules22091430
Ramón-García S, Mikut R, Ng C, Ruden S, Volkmer R, Reischl M, Hilpert K, Thompson CJ (2013) Targeting Mycobacterium tuberculosis and other microbial pathogens using improved synthetic antibacterial peptides. Antimicrob Agents Chemother 57(5):2295–2303. https://doi.org/10.1128/AAC.00175-13
Santos P, Gordillo A, Osses L, Salazar LM, Soto CY (2012) Effect of antimicrobial peptides on ATPase activity and proton pumping in plasma membrane vesicles obtained from mycobacteria. Peptides 36(1):121–128. https://doi.org/10.1016/j.peptides.2012.04.018
Sieniawska E, Sawicki R, Golus J, Swatko-Ossor M, Ginalska G, Skalicka-Wozniak K (2018) Nigella damascena L. essential oil—a valuable source of β-elemene for antimicrobial testing. Molecules 23(2):256. https://doi.org/10.3390/molecules23020256
Sirgel FA, Wiid IJ, van Helden PD (2009) Measuring minimum inhibitory concentrations in mycobacteria. In: Parish T, Brown AC (eds) Mycobacteria protocols. Humana Press, Totowa, pp 173–186
von Groll A, Martin A, Portaels F, da Silva PE, Palomino JC (2010) Growth kinetics of Mycobacterium tuberculosis measured by quantitative resazurin reduction assay: a tool for fitness studies. Braz J Microbiol 41(2):300–303. https://doi.org/10.1590/S1517-83822010000200006
Webster AM, Cobb SL (2018) Recent advances in the synthesis of peptoid macrocycles. Chemistry 24(30):7560–7573. https://doi.org/10.1002/chem.201705340
Zuckermann RN, Kerr JM, Kent SBH, Moos WH (1992) Efficient method for the preparation of peptoids oligo(N-substituted glycines) by submonomer solid-phase synthesis. J Am Chem Soc 114(26):10646–10647. https://doi.org/10.1021/ja00052a076
Acknowledgements
Gopal Singh Bisht would like to acknowledge Indian Council for Medical Research, Delhi for providing financial support (Project ID 52/4/2013-Bio BMS) and Central Drug Research Institute, Lucknow for providing bacterial strain M. smegmatis MC2155.
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All procedures and protocols mentioned in this article have been approved by Institutional Biosafety Committee of Jaypee University of Information Technology, Waknaghat. The study does not involve any human participant or animal.
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Sharma, D., Poonam, Shrivastava, R. et al. In Vitro Efficacy of Lipid Conjugated Peptidomimetics Against Mycobacterium smegmatis. Int J Pept Res Ther 26, 531–537 (2020). https://doi.org/10.1007/s10989-019-09859-7
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DOI: https://doi.org/10.1007/s10989-019-09859-7