High-resolution structural profile of hylaseptin-4: Aggregation, membrane topology and pH dependence of overall membrane binding process

• Published in: Biochimica et biophysica acta. Biomembranes Vol. 1863; no. 5; p. 183581
• Main Authors: Nunes, L.O., Munhoz, V.H.O., Sousa, A.A., de Souza, K.R., Santos, T.L., Bemquerer, M.P., Ferreira, D.E.C., de Magalhães, Mariana T.Q., Resende, J.M., Alcântara, A.F.C., Aisenbrey, C., Veloso, D.P., Bechinger, B., Verly, R.M.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.05.2021; Elsevier
• Subjects: Aggregation process; Antimicrobial mechanism of action; Biophysical prediction of peptidemembrane interactions; Chemical Sciences; Conformational analysis of peptides; Membrane active peptides; Other; Peptide-membrane interaction; Peptide-membrane interaction; Aggregation process; Biophysical prediction of peptidemembrane interactions; Antimicrobial mechanism of action; Conformational analysis of peptides; Membrane active peptides
• ISSN: 0005-2736; 1879-2642
• DOI: 10.1016/j.bbamem.2021.183581

Hylaseptin-4 (HSP-4, GIGDILKNLAKAAGKAALHAVGESL-NH2) is an antimicrobial peptide originally isolated from Hypsiboas punctatus tree frog. The peptide has been chemically synthetized for structural investigations by CD and NMR spectroscopies. CD experiments reveal the high helical content of HSP-4 in biomimetic media. Interestingly, the aggregation process seems to occur at high peptide concentrations either in aqueous solution or in presence of biomimetic membranes, indicating an increase in the propensity of the peptide for adopting a helical conformation. High-resolution NMR structures determined in presence of DPC-d38 micelles show a highly ordered α-helix from amino acid residues I2 to S24 and a smooth bend near G14. A large separation between hydrophobic and hydrophilic residues occurs up to the A16 residue, from which a shift in the amphipathicity is noticed. Oriented solid-state NMR spectroscopy show a roughly parallel orientation of the helical structure along the POPC lipid bilayer surface, with an insertion of the hydrophobic N-terminus into the bilayer core. Moreover, a noticeable pH dependence of the aggregation process in both aqueous and in biomimetic membrane environments is attributed to a single histidine residue (H19). The protonation degree of the imidazole side-chain might help in modulating the peptide-peptide or peptide-lipid interactions. Finally, molecular dynamics simulations confirm the orientation and preferential helical conformation and in addition, show that HSP-4 tends to self-aggregate in order to stabilize its active conformation in aqueous or phospholipid bilayer environments. [Display omitted] •Concentration and pH-dependent peptide binding to the biomembranes.•High-resolution structure and orientation of hylaseptin-4 in biomimetic environments•Amphipathic phase shift plays an important role on the self-aggregation process near the membrane interface