Skip to main content
U.S. flag

An official website of the United States government

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you’ve safely connected to the .gov website. Share sensitive information only on official, secure websites.

Structure and Function in Antimicrobial Piscidins: Histidine Position, Directionality of Membrane Insertion, and pH-Dependent Permeabilization

Published

Author(s)

Mihaela Mihailescu, Mirco Sorci, Jolita Seckute, Vitalii I. Silin, Janet Hammer, B. Scott Perrin, Jorge Hernandez, Nedzada Smajic, Akritee Shrestha, Kimberly Bogadardus, Alexander Greenwood, Riqiang Fu, Jack Blazyk, Richard W. Pastor, Linda Nicholson, Georges Belfort, Myriam Cotten

Abstract

Piscidins are histidine-enriched antimicrobial peptides that interact with lipid bilayers as amphipathic alpha-helices. Their activity at acidic and basic pH in vivo makes them promising templates for biomedical applications. This study focuses on p1 and p3, both 22-residue-long piscidins with 68% sequence identity. They share three histidines (H3, H4, and H11), but p1, which is significantly more permeabilizing, has a fourth histidine (H17). This study investigates how variations in amphipathic character associated with histidines affect the permeabilization properties of p1 and p3. First, we show that the permeabilization ability of p3, but not p1, is strongly inhibited at pH 6.0 when the conserved histidines are partially charged and H17 is predominantly neutral. Second, our neutron diffraction measurements performed at low water content and neutral pH indicate that the average conformation of p1 is highly tilted, with its C-terminus extending into the opposite leaflet. In contrast, p3 is surface bound with its N-terminal end tilted toward the bilayer interior. The deeper membrane insertion of p1 correlates with its behavior at full hydration: an enhanced ability to tilt, bury its histidines and C-terminus, induce membrane thinning and defects, and alter membrane conductance and viscoelastic properties. Furthermore, its pH-resiliency relates to the neutral state favored by H17. Overall, these results provide mechanistic insights into how differences in the histidine content and amphipathicity of peptides can elicit different directionality of membrane insertion and pH-dependent permeabilization. This work features complementary methods, including dye leakage assays, NMR-monitored titrations, X-ray and neutron diffraction, oriented CD, molecular dynamics, electrochemical impedance spectroscopy, surface plasmon resonance, and quartz crystal microbalance with dissipation.
Citation
Journal of the American Chemical Society
Volume
141
Issue
25

Keywords

SOLID-STATE NMR, HOST-DEFENSE PEPTIDES, LIPID-BILAYERS, X-RAY, MOLECULAR RECOGNITION, NEUTRON-DIFFRACTION, MODEL MEMBRANES, FORCE-FIELD, MAST-CELLS, MECHANISM

Citation

Mihailescu, M. , Sorci, M. , Seckute, J. , Silin, V. , Hammer, J. , Perrin, B. , Hernandez, J. , Smajic, N. , Shrestha, A. , Bogadardus, K. , Greenwood, A. , Fu, R. , Blazyk, J. , Pastor, R. , Nicholson, L. , Belfort, G. and Cotten, M. (2019), Structure and Function in Antimicrobial Piscidins: Histidine Position, Directionality of Membrane Insertion, and pH-Dependent Permeabilization, Journal of the American Chemical Society, [online], https://doi.org/10.1021/jacs.9b00440 (Accessed April 24, 2024)
Created May 30, 2019, Updated October 12, 2022