Influence of disulfide bonds in human beta defensin-3 on its strain specific activity against Gram-negative bacteria

Nehls, Christian; Böhling, Arne; Podschun, Rainer; Schubert, Sabine; Grötzinger, Joachim; Schromm, Andra B.; Fedders, Henning; Leippe, Matthias; Harder, Jürgen; Kaconis, Yani; Gronow, Sabine; Gutsmann, Thomas

doi:10.1016/j.bbamem.2020.183273

Cited by 18 publications

(17 citation statements)

References 53 publications

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“…This indicates that the antibacterial effect of the AMPs designed by us is stronger than that of most studied AMPs. Among our AMPs, S-6 with added pheromone, enhanced the antibacterial effect against the Gram-negative bacteria P. mendocina and V. parahaemolyticus , which may be because of the fact that the peptide chain contains a pair of disulfide bonds, which can increase the stability of AMPs and improve bioavailability ( Nehls et al, 2020 ). Among the AMPs, P-6, S-6, and L-6, with the addition of pheromones, showed enhanced bacteriostatic effects against Gram-negative bacteria P. mendocina , and P-6 and S-6 showed enhanced bacteriostatic effects against Gram-negative bacteria V. parahaemolyticus , which may be attributed to the existence of Lipopolysaccharides (LPS) on the cell membranes of Gram-negative bacteria.…”

Section: Discussionmentioning

confidence: 99%

Improving the Activity of Antimicrobial Peptides Against Aquatic Pathogen Bacteria by Amino Acid Substitutions and Changing the Ratio of Hydrophobic Residues

Tan

Wang

et al. 2021

Front. Microbiol.

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With the increasing number of drug-resistant bacteria, there is an urgent need for new antimicrobial agents, and antimicrobial peptides (AMPs), which exist in the human non-specific immune system, are one of the most promising candidates. It is an effective optimization strategy to modify antimicrobial peptides (AMPs) according to the distribution of amino acids and hydrophobic characteristics. The addition of bacterial pheromones to the N short peptide can increase the ability to recognize bacteria. In this study, we designed and synthesized AMP1–6 by amino acid substitution of mBjAMP1. Additionally, P-6, S-6, and L-6 were designed and synthesized by adding bacterial pheromones based on 1–6. Functional tests showed that the four AMPs had the ability to kill Gram-negative Vibrio anguillarum, Pseudomonas mendocina, and Vibrio parahaemolyticus, and Gram-positive Micrococcus luteus and Listeria monocytogenes. Additionally, all four AMPs induced permeabilization and depolarization of bacterial cell membranes and increased intracellular reactive oxygen species (ROS) levels. Importantly, they had little or no mammalian cytotoxicity. At the same time, 1–6 and L-6 protected the stability of intestinal flora in Sebastes schlegelii and increased the relative abundance of Lactobacillaceae. In summary, our results indicate that the designed AMPs have broad application prospects as a new type of polypeptide antimicrobial agent.

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Section: Discussionmentioning

confidence: 99%

Improving the Activity of Antimicrobial Peptides Against Aquatic Pathogen Bacteria by Amino Acid Substitutions and Changing the Ratio of Hydrophobic Residues

Tan

Wang

et al. 2021

Front. Microbiol.

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“…There are many articles about the mechanism of human β -defensins killing bacteria. Most defensins kill bacteria by destroying the bacterial biofilm structure of bacteria [ 37 , 38 ]. In the previous study of DEFB118, we found that DEFB118 destroyed the normal morphology and structure of bacteria by increasing the permeability of the bacterial membrane, causing bacterial death [ 36 ].…”

Section: Discussionmentioning

confidence: 99%

Expression and Functional Characterization of a Novel Antimicrobial Peptide: Human Beta-Defensin 118

Lin

Xie

Chen

et al. 2020

BioMed Research International

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Purpose. β-Defensin 118 (DEFB118) is a novel host defense peptide (HDP) identified in humans. To evaluate its potentials for future utilization, the DEFB118 gene was expressed in Escherichia coli (E. coli) and the recombinant protein was fully characterized. Methods. The DEFB118 protein was obtained by heterologous expression using E. coli Rosetta (DE3). Antibacterial activity of DEFB118 was determined by using various bacterial strains. IPEC-J cells challenged by E. coli K88 were used to determine its influences on inflammatory responses. Results. The E. coli transformants yielded more than 250 μg/mL DEFB118 protein after 4 h induction by 1.0 mM IPTG. The DEFB118 was estimated by SDS-PAGE to be 30 kDa, and MALDI-TOF analysis verified that it is a human β-defensin 118. Importantly, the DEFB118 showed antimicrobial activities against both Gram-negative bacteria (E. coli K88 and E. coli DH5α) and Gram-positive bacteria (S. aureus and B. subtilis), with a minimum inhibitory concentration (MIC) of 4 μg/mL. Hemolytic assays showed that DEFB118 had no detrimental impact on cell viability. Additionally, DEFB118 was found to elevate the viability of IPEC-J2 cells upon E. coli K88 challenge. Moreover, DEFB118 significantly decreased cell apoptosis in the late apoptosis phase and downregulated the expression of inflammatory cytokines such as IL-1β and TNF-α in IPEC-J2 cell exposure to E. coli K88. Conclusions. These results suggested a novel function of the mammalian defensins, and the antibacterial and anti-inflammatory properties of DEFB118 may allow it as a potential substitute for conventionally used antibiotics or drugs.

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“…The linear hβD-3 variant was synthesized and purified as previously described. 34 The lyophilized peptides to be tested were freshly hydrated in Milli-Q water briefly before the experiment except for Alamethicin due to its poor solubility in water, the Alamethicin stock was dissolved in Ethanol. Aliquots were then diluted to the desired peptide concentrations in the same solution as the IA phase; a small amount of HPTS (5 μM) was also added as a tracer to precisely time peptide arrival in the GUV trap chambers.…”

Section: Methodsmentioning

confidence: 99%

“…Polymyxin B lyses membranes in a detergent-like manner, 24 while the mechanism of the linear form of hβD-3 is believed to involve disordered toroidal pore formation. 34 The concentration at which these peptides were studied, the lipid composition of reconstituted membranes used in the studies, and other experimental parameters are factors that may affect their activity characteristics. With this in mind, we characterized the modes of action of these peptides using the GUV studio under the conditions used for the first series, to probe unique signatures for the LE time distributions for these peptides.…”

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confidence: 99%

Measuring thousands of single vesicle leakage events reveals the mode of action of antimicrobial peptides

Nahas

Fletcher

Hammond

et al. 2021

Preprint

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Host defense or antimicrobial peptides hold promise for providing new pipelines of effective antimicrobial agents. Their activity quantified against model phospholipid membranes is fundamental to a detailed understanding of their structure-activity relationships. However, existing characterization assays lack the resolution necessary to achieve this insight. Leveraging a highly parallelized microfluidic platform for trapping and studying thousands of giant unilamellar vesicles, we conducted quantitative long-term microscopy studies to monitor the membrane-disruptive activity of archetypal antimicrobial peptides with a high spatiotemporal resolution. We described the modes of action of these peptides via measurements of the disruption of the vesicle population under the conditions of continuous peptide dosing using a range of concentrations, and related the observed modes with the molecular activity mechanisms of these peptides. The study offers an effective approach for characterizing membrane-targeting antimicrobial agents in a standardized manner, and for assigning specific modes of action to the corresponding antimicrobial mechanisms.

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Influence of disulfide bonds in human beta defensin-3 on its strain specific activity against Gram-negative bacteria

Cited by 18 publications

References 53 publications

Improving the Activity of Antimicrobial Peptides Against Aquatic Pathogen Bacteria by Amino Acid Substitutions and Changing the Ratio of Hydrophobic Residues

Improving the Activity of Antimicrobial Peptides Against Aquatic Pathogen Bacteria by Amino Acid Substitutions and Changing the Ratio of Hydrophobic Residues

Expression and Functional Characterization of a Novel Antimicrobial Peptide: Human Beta-Defensin 118

Measuring thousands of single vesicle leakage events reveals the mode of action of antimicrobial peptides

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