Antimicrobial peptide RP‐1 structure and interactions with anionic versus zwitterionic micelles

Bourbigot, Sarah; Dodd, Erin L.; Horwood, Chrystal; Cumby, Nichole; Fardy, Liam; Welch, William H.; Ramjan, Zachary; Sharma, Sanjai; Waring, Alan J.; Yeaman, Michael R.; Booth, Valerie

doi:10.1002/bip.21071

Cited by 51 publications

(67 citation statements)

References 65 publications

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“…The similarity of TRP3 conformations and of some of its analogs stabilized by different micellar interfaces has been indicated by NMR, CD, and fluorescence experiments (13,14,17,29,70). Similar results were found for melittin in micelles of various detergentes (45) and for PR-1, a synthetic peptide modeled after the microbicide domains of the platelet microparticle factor-4 family, in SDS and DPC micelles (71).…”

Section: Conformation Of Trp3 Bound To the Micellar Systemssupporting

confidence: 62%

Structural and Dynamic Insights of the Interaction between Tritrpticin and Micelles: An NMR Study

Santos

Moraes

Nakaie

et al. 2016

Biophysical Journal

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A large number of antimicrobial peptides (AMPs) acts with high selectivity and specificity through interactions with membrane lipid components. These peptides undergo complex conformational changes in solution; upon binding to an interface, one major conformation is stabilized. Here we describe a study of the interaction between tritrpticin (TRP3), a cathelicidin AMP, and micelles of different chemical composition. The peptide's structure and dynamics were examined using one-dimensional and two-dimensional NMR. Our data showed that the interaction occurred by conformational selection and the peptide acquired similar structures in all systems studied, despite differences in detergent headgroup charge or dipole orientation. Fluorescence and paramagnetic relaxation enhancement experiments showed that the peptide is located in the interface region and is slightly more deeply inserted in 1-myristoyl-2-hydroxy-sn-glycero-3-phospho-1 0 -rac-glycerol (LMPG, anionic) than in 1-lauroyl-2-hydroxy-sn-glycero-3-phosphocholine (LLPC, zwitterionic) micelles. Moreover, the tilt angle of an assumed helical portion of the peptide is similar in both systems. In previous work we proposed that TRP3 acts by a toroidal pore mechanism. In view of the high hydrophobic core exposure, hydration, and curvature presented by micelles, the conformation of TRP3 in these systems could be related to the peptide's conformation in the toroidal pore.

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Section: Conformation Of Trp3 Bound To the Micellar Systemssupporting

confidence: 62%

Structural and Dynamic Insights of the Interaction between Tritrpticin and Micelles: An NMR Study

Santos

Moraes

Nakaie

et al. 2016

Biophysical Journal

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“…The synthetic host defense peptide RP-1 is an 18-amino-acid peptide (N-ALYKK 5 -FKKKL 10 -LKSLK 15 -RLG 18 -C; mass, 2,163 Da) modeled in part upon ␣-helical C-terminal microbicidal domains of mammalian CXCL4 kinocidins (5). AA-RP-1 is an anthryl-alanine-substituted (position 2) congener of the parent peptide RP-1.…”

Section: Methodsmentioning

confidence: 99%

“…AA-RP-1 is an anthryl-alanine-substituted (position 2) congener of the parent peptide RP-1. Peptides were synthesized with a Symphony multiplex synthesizer (Rainin, Woburn, MA) and authenticated by mass spectroscopy and amino acid analysis (5,46). Purified peptides were lyophilized and resuspended in sterile phosphate-buffered saline (PBS) for use in the assays.…”

Section: Methodsmentioning

confidence: 99%

“…RP-1 and congener peptides such as AA-RP-1 exert potent microbicidal efficacy in complex blood and blood-derived matrices, with minimal cytotoxicity against erythrocytes and endothelial cells (5,46). We describe here the antiprotozoan activity of RP-1 and its analogue AA-RP-1 evaluated for specific leishmanicidal activity, mechanisms of action, and efficacy in a mouse model of experimental visceral leishmaniasis.…”

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confidence: 99%

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Efficacy of Synthetic Peptides RP-1 and AA-RP-1 against Leishmania Species In Vitro and In Vivo

Erfe

David

Huang

et al. 2012

Antimicrob Agents Chemother

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Host defense peptides are naturally occurring molecules that play essential roles in innate immunity to infection. Based on prior structure-function knowledge, we tested two synthetic peptides (RP-1 and AA-RP-1) modeled on the conserved, microbicidal ␣-helical domain of mammalian CXCL4 platelet kinocidins. These peptides were evaluated for efficacy against Leishmania species, the causative agents of the group of diseases known as leishmaniasis. In vitro antileishmanial activity was assessed against three distinct Leishmania strains by measuring proliferation, metabolic activity and parasite viability after exposure to various concentrations of peptides. We demonstrate that micromolar concentrations of RP-1 and AA-RP-1 caused dose-dependent growth inhibition of Leishmania promastigotes. This antileishmanial activity correlated with rapid membrane disruption, as well as with a loss of mitochondrial transmembrane potential. In addition, RP-1 and AA-RP-1 demonstrated distinct and significant in vivo antileishmanial activities in a mouse model of experimental visceral leishmaniasis after intravenous administration. These results establish efficacy of RP-1 lineage synthetic peptides against Leishmania species in vitro and after intravenous administration in vivo and provide further validation of proof of concept for the development of these and related systemic antiinfective peptides targeting pathogens that are resistant to conventional antibiotics.

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“…Detailed structure-function studies have shown that positively charged residues on the surface of these chemokines, especially at their C-terminal ␣-helices, are largely responsible for their direct antimicrobial activity (4,9,28,39,56,63). Peptides of the ␣-helical portions of some chemokines have been shown to have preferential interactions with anionic lipids over zwitterionic lipids (5,6), and the full chemokines are believed to act by perturbing bacterial membranes similarly to antimicrobial peptides (10,17).…”

mentioning

confidence: 99%

Exploring Platelet Chemokine Antimicrobial Activity: Nuclear Magnetic Resonance Backbone Dynamics of NAP-2 and TC-1

Nguyen

Kwakman

Chan

et al. 2011

Antimicrob Agents Chemother

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The platelet chemokines neutrophil-activating peptide-2 (NAP-2) and thrombocidin-1 (TC-1) differ by only two amino acids at their carboxy-terminal ends. Nevertheless, they display a significant difference in their direct antimicrobial activities, with the longer NAP-2 being inactive and TC-1 being active. In an attempt to rationalize this difference in activity, we studied the structure and the dynamics of both proteins by nuclear magnetic resonance (NMR) spectroscopy. Using 15 N isotope-labeled protein, we confirmed that the two monomeric proteins essentially have the same overall structure in aqueous solution. However, NMR relaxation measurements provided evidence that the negatively charged carboxy-terminal residues of NAP-2 experience a restricted motion, whereas the carboxy-terminal end of TC-1 moves in an unrestricted manner. The same behavior was also seen in molecular dynamic simulations of both proteins. Detailed analysis of the protein motions through model-free analysis, as well as a determination of their overall correlation times, provided evidence for the existence of a monomer-dimer equilibrium in solution, which seemed to be more prevalent for TC-1. This finding was supported by diffusion NMR experiments. Dimerization generates a larger cationic surface area that would increase the antimicrobial activities of these chemokines. Moreover, these data also show that the negatively charged carboxy-terminal end of NAP-2 (which is absent in TC-1) folds back over part of the positively charged helical region of the protein and, in doing so, interferes with the direct antimicrobial activity.

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Antimicrobial peptide RP‐1 structure and interactions with anionic versus zwitterionic micelles

Cited by 51 publications

References 65 publications

Structural and Dynamic Insights of the Interaction between Tritrpticin and Micelles: An NMR Study

Structural and Dynamic Insights of the Interaction between Tritrpticin and Micelles: An NMR Study

Efficacy of Synthetic Peptides RP-1 and AA-RP-1 against Leishmania Species In Vitro and In Vivo

Exploring Platelet Chemokine Antimicrobial Activity: Nuclear Magnetic Resonance Backbone Dynamics of NAP-2 and TC-1

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