Version 1
: Received: 29 August 2024 / Approved: 29 August 2024 / Online: 30 August 2024 (03:37:03 CEST)
How to cite:
Huang, S.; Su, G.; Yang, L.; Yue, L.; Chen, L.; Huang, J.; Yang, F. Single−molecule Level Quantification Reveals the Interaction between Melittin and Lipopolysaccharide by Atomic Force Microscopy. Preprints2024, 2024082140. https://doi.org/10.20944/preprints202408.2140.v1
Huang, S.; Su, G.; Yang, L.; Yue, L.; Chen, L.; Huang, J.; Yang, F. Single−molecule Level Quantification Reveals the Interaction between Melittin and Lipopolysaccharide by Atomic Force Microscopy. Preprints 2024, 2024082140. https://doi.org/10.20944/preprints202408.2140.v1
Huang, S.; Su, G.; Yang, L.; Yue, L.; Chen, L.; Huang, J.; Yang, F. Single−molecule Level Quantification Reveals the Interaction between Melittin and Lipopolysaccharide by Atomic Force Microscopy. Preprints2024, 2024082140. https://doi.org/10.20944/preprints202408.2140.v1
APA Style
Huang, S., Su, G., Yang, L., Yue, L., Chen, L., Huang, J., & Yang, F. (2024). Single−molecule Level Quantification Reveals the Interaction between Melittin and Lipopolysaccharide by Atomic Force Microscopy. Preprints. https://doi.org/10.20944/preprints202408.2140.v1
Chicago/Turabian Style
Huang, S., Jinxiu Huang and Feiyun Yang. 2024 "Single−molecule Level Quantification Reveals the Interaction between Melittin and Lipopolysaccharide by Atomic Force Microscopy" Preprints. https://doi.org/10.20944/preprints202408.2140.v1
Abstract
The interaction forces and mechanical properties of the interaction between melittin (Mel) and lipopolysaccharide (LPS) are considered a crucial driving force for Mel in killing Gram−negative bacteria (GNB). However, how their interaction forces are performed at the single−molecule level and the dissociation kinetic characteristics of the Mel/LPS complex remain poorly understood. In this study, the single−molecule level interaction forces between Mel and LPSs from E. coil K−12, O55:B5, O111:B4, and O128:B12 were explored using atomic force microscopy (AFM) based single−molecule force spectroscopy (SMFS). Further, the AFM−based dynamic force spectroscopy (DFS) and advanced analytical model were employed to investigate the kinetic characteristics of the Mel/LPSs complex dissociation. The results indicated that Mel could interact with both rough (R)−form LPS (E. coli K−12) and smooth (S)−form LPSs (E. coli O55:B5, O111:B4, O128:B12), and the S−form LPSs showed a more robust interaction with Mel than R−form LPS, and a slight difference existed in the interaction forces between Mel and diverse S−form LPSs. The Mel interacts with S−form LPSs showed greater specific and non−specific interaction forces than R−form LPS (p < 0.05), as determined by AFM−based SMFS. However, there was no significant difference in the specific and non−specific interaction forces among the three S−form LPSs (p > 0.05), indicating the variability of O−antigen could not affect the interaction between Mel and LPSs. The DFS result showed that the Mel/S−form LPSs complexes had a lower dissociation rate constant (koff), a shorter energy barrier width (xβ), a longer bond lifetime (τoff), and a higher height of the energy barrier (∆G), demonstrating that Mel could interact with S−form LPSs to form more stable complexes. In conclusion, this study promotes our knowledge of the interaction micromechanics and kinetics characteristics between Mel and LPS at the single−molecule level. Moreover, it can help us design and evaluate new anti−GNB drugs.
Keywords
melittin; lipopolysaccharide; atomic force microscopy; interaction force; kinetics characteristics
Subject
Biology and Life Sciences, Biochemistry and Molecular Biology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
