[Antibacterial Mechanism of Brevinin-2GHk, an Antimicrobial Peptide from Skin]

Overview

Journal Nan Fang Yi Ke Da Xue Xue Bao

Specialty General Medicine

Date 2021 Dec 17

PMID 34916191

Citations 1

Authors

W Xiong

J Chai

J Wu

M Tian

W Lu

X Xu

Affiliations

Soon will be listed here.

Abstract

Objective: To investigate the secondary structure, physicochemical properties and antibacterial activity of Brevinin- 2GHk (BR2GK), an antimicrobial peptide from frog skin, and explore its antibacterial mechanism.

Methods: BR2GK was synthesized, purified with high performance liquid chromatography (HPLC) and identified using mass spectrometry. Circular dichroism was used to determine the secondary structure and physicochemical properties of BR2GK. Two-fold dilution method was used to determine the antibacterial activity of BR2GK, and its antibacterial mechanism was explored using laser scanning confocal microscopy (LSCM) and scanning electron microscopy (SEM). The hemolytic activity of BR2GK was analyzed in red blood cells. Isothermal titration calorimetry (ITC) and surface plasmon resonance imaging (SPRi) were employed to detect the binding of BR2GK to lipopolysaccharide (LPS), and the antioxidant activity of BR2GK was evaluated using biochemical kits.

Results: The synthesized BR2GK, with a purity exceeding 95% after purification, had the basic characteristics of cationic antimicrobial peptides. BR2GK consisted mainly of α-helical structure in SDS solution and exhibited a broad-spectrum antibacterial activity. Antibacterial activity assay showed that BR2GK had inhibitory and killing activity against a variety of strains with a minimum inhibitory concentration (MIC) of 2.76 μmol/L against . Observation with LSCM and SEM showed that BR2GK at an active concentration caused bacterial cell membrane damage, cell swelling, contraction, deformation, and massive exudation of intracellular contents without causing hemolysis. ITC showed that the binding affinity K of BR2GK to LPS was 18.2±0.8 μmol/L. The antioxidant test showed that BR2GK was capable of effectively scavenging NO, ABTS and DPPH.

Conclusion: BR2GK, as a broad-spectrum antibacterial peptide with also a strong antioxidant capacity, does not cause hemolysis and is capable of binding to LPS. BR2GK has an important value for future design and synthesis of antimicrobial peptides with stronger antibacterial activity and lower cytotoxicity.

Citing Articles

Assessment of New Strategies to Improve the Performance of Antimicrobial Peptides.

Wang L, Liu H, Li X, Yao C Nanomaterials (Basel). 2022; 12(20).

PMID: 36296881 PMC: 9610275. DOI: 10.3390/nano12203691.

References

Lin Y, Liu S, Xi X, Ma C, Wang L, Chen X . Study on the Structure-Activity Relationship of an Antimicrobial Peptide, Brevinin-2GUb, from the Skin Secretion of . Antibiotics (Basel). 2021; 10(8). PMC: 8388802. DOI: 10.3390/antibiotics10080895. View

Peng J, Yang Y, Zhao P, Qiu S, Jia F, Wang J . Cu2+ reduces hemolytic activity of the antimicrobial peptide HMPI and enhances its trypsin resistance. Acta Biochim Biophys Sin (Shanghai). 2020; 52(6):603-611. DOI: 10.1093/abbs/gmaa043. View

Huang C, Wang S, Chen I, Chen Y, Liu P, Fang S . Protective Effect of Piplartine against LPS-Induced Sepsis through Attenuating the MAPKs/NF-κB Signaling Pathway and NLRP3 Inflammasome Activation. Pharmaceuticals (Basel). 2021; 14(6). PMC: 8234963. DOI: 10.3390/ph14060588. View

Zeng B, Chai J, Deng Z, Ye T, Chen W, Li D . Functional Characterization of a Novel Lipopolysaccharide-Binding Antimicrobial and Anti-Inflammatory Peptide in Vitro and in Vivo. J Med Chem. 2018; 61(23):10709-10723. DOI: 10.1021/acs.jmedchem.8b01358. View

Som A, Vemparala S, Ivanov I, Tew G . Synthetic mimics of antimicrobial peptides. Biopolymers. 2008; 90(2):83-93. DOI: 10.1002/bip.20970. View

Greco I, Molchanova N, Holmedal E, Jenssen H, Hummel B, Watts J . Correlation between hemolytic activity, cytotoxicity and systemic in vivo toxicity of synthetic antimicrobial peptides. Sci Rep. 2020; 10(1):13206. PMC: 7414031. DOI: 10.1038/s41598-020-69995-9. View

Chen Y, Guarnieri M, Vasil A, Vasil M, Mant C, Hodges R . Role of peptide hydrophobicity in the mechanism of action of alpha-helical antimicrobial peptides. Antimicrob Agents Chemother. 2006; 51(4):1398-406. PMC: 1855469. DOI: 10.1128/AAC.00925-06. View

Mahlapuu M, Hakansson J, Ringstad L, Bjorn C . Antimicrobial Peptides: An Emerging Category of Therapeutic Agents. Front Cell Infect Microbiol. 2017; 6:194. PMC: 5186781. DOI: 10.3389/fcimb.2016.00194. View

Huan Y, Kong Q, Mou H, Yi H . Antimicrobial Peptides: Classification, Design, Application and Research Progress in Multiple Fields. Front Microbiol. 2020; 11:582779. PMC: 7596191. DOI: 10.3389/fmicb.2020.582779. View

10.

Murtaza G, Rizvi A, Irfan M, Yan D, Khan R, Rafique B . Glycated albumin based photonic crystal sensors for detection of lipopolysaccharides and discrimination of Gram-negative bacteria. Anal Chim Acta. 2020; 1117:1-8. DOI: 10.1016/j.aca.2020.04.018. View

11.

Chen G, Miao Y, Ma C, Zhou M, Shi Z, Chen X . Brevinin-2GHk from and the Design of Truncated Analogs Exhibiting the Enhancement of Antimicrobial Activity. Antibiotics (Basel). 2020; 9(2). PMC: 7168151. DOI: 10.3390/antibiotics9020085. View

12.

Pei X, Gong Z, Wu Q, Chen X, Wang L, Ma C . Characterisation of a novel peptide, Brevinin-1H, from the skin secretion of Amolops hainanensis and rational design of several analogues. Chem Biol Drug Des. 2020; 97(2):273-282. DOI: 10.1111/cbdd.13779. View

13.

Wang X, Ren S, Guo C, Zhang W, Zhang X, Zhang B . Identification and functional analyses of novel antioxidant peptides and antimicrobial peptides from skin secretions of four East Asian frog species. Acta Biochim Biophys Sin (Shanghai). 2017; 49(6):550-559. DOI: 10.1093/abbs/gmx032. View

14.

Higgins M, Eswaran J, Edwards P, Schertler G, Hughes C, Koronakis V . Structure of the ligand-blocked periplasmic entrance of the bacterial multidrug efflux protein TolC. J Mol Biol. 2004; 342(3):697-702. DOI: 10.1016/j.jmb.2004.07.088. View

15.

Kumari P, Russo A, Wright S, Muthupalani S, Rathinam V . Hierarchical cell-type-specific functions of caspase-11 in LPS shock and antibacterial host defense. Cell Rep. 2021; 35(3):109012. PMC: 8451177. DOI: 10.1016/j.celrep.2021.109012. View

16.

Madanchi H, Ebrahimi Kiasari R, Seyed Mousavi S, Johari B, Shabani A, Sardari S . Design and Synthesis of Lipopolysaccharide-Binding Antimicrobial Peptides Based on Truncated Rabbit and Human CAP18 Peptides and Evaluation of Their Action Mechanism. Probiotics Antimicrob Proteins. 2020; 12(4):1582-1593. DOI: 10.1007/s12602-020-09648-5. View

17.

Fujiwara Y, Ohnishi K, Horlad H, Saito Y, Shiraishi D, Takeya H . CD163 deficiency facilitates lipopolysaccharide-induced inflammatory responses and endotoxin shock in mice. Clin Transl Immunology. 2020; 9(9):e1162. PMC: 7518957. DOI: 10.1002/cti2.1162. View

18.

Xu X, Lai R . The chemistry and biological activities of peptides from amphibian skin secretions. Chem Rev. 2015; 115(4):1760-846. DOI: 10.1021/cr4006704. View

19.

Maria-Neto S, de Almeida K, Macedo M, Franco O . Understanding bacterial resistance to antimicrobial peptides: From the surface to deep inside. Biochim Biophys Acta. 2015; 1848(11 Pt B):3078-88. DOI: 10.1016/j.bbamem.2015.02.017. View

20.

Zhang Y, Skolnick J . TM-align: a protein structure alignment algorithm based on the TM-score. Nucleic Acids Res. 2005; 33(7):2302-9. PMC: 1084323. DOI: 10.1093/nar/gki524. View