Journal Article

Design and Synthesis of Lipopolysaccharide-Binding Antimicrobial Peptides Based on Truncated Rabbit and Human CAP18 Peptides and Evaluation of Their Action Mechanism

Seyed Mousavi, Seyed Javad, Johari, Behrooz, Madanchi, Hamid, Ebrahimi Kiasari, Ramin, Sardari, Soroush, Shabani, Ali Akbar

2020

Overview

Lipopolysaccharide (LPS) is a toxic and immunogenic agent for human. Additionally, LPS is a good target for some antimicrobial compounds, including antimicrobial peptides (AMPs). LPS-binding peptides (LBPs) can recognize and neutralize LPS. Rabbit and human cathelicidins are AMPs with LPS-binding activity. In this study, we designed and synthesized two new truncated LBPs from rabbit and human CAP18 peptides by in silico methods. After synthesis of peptides, the antimicrobial properties and LPS-binding activity of these peptides were evaluated. The parental rabbit and human CAP18 peptides were selected as positive controls. Next, the changes in the secondary structure of these peptides before and after treatment with LPS were measured by circular dichroism (CD). Human cytotoxicity of the peptides was evaluated by MTT and red blood cells (RBCs) hemolysis assays. Finally, field emission scanning electron microscopy (FE-SEM), confocal microscopy, and flow cytometry were performed to study the action mechanism of these peptides. Results indicated that the hCap18 and rCap18 had antibacterial activity (at a MIC of 4–128 μg/mL). The results of the quantitative LAL test demonstrated that LPS-binding activity of hCap18 peptide was better than rCap18, while rCap18 peptide had better antimicrobial properties. Furthermore, rCap18 had less cytotoxicity than hCap18. However, both peptides were nontoxic for normal human skin fibroblast cell in MIC range. In conclusion, rCap18 has good antibacterial properties, while hCap18 can be tested as a diagnostic molecule in our future studies.

• Publisher: Springer US,Springer Nature B.V
• Subject: Antibacterial activity / antibacterial properties / Antimicrobial peptides / Applied Microbiology / Cathelicidins / Chemistry / Chemistry and Materials Science / Chemistry/Food Science / Circular dichroism / circular dichroism spectroscopy / computer simulation / Confocal microscopy / Cytotoxicity / Erythrocytes / fibroblasts / Flow cytometry / hemolysis / humans / Immunogenicity / Limulus amebocyte lysate assay / Lipopolysaccharides / Microbiology / Microscopy / Minimum inhibitory concentration / Nutrition / Peptides / probiotics / Protein Science / Protein structure / rabbits / Scanning electron microscopy / Secondary structure / skin (animal)