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

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

doi:10.1021/jacs.9b00440

Cited by 64 publications

(123 citation statements)

References 122 publications

(282 reference statements)

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“…The most N-terminal Arg residue, Arg5, could become static because the N-terminal His-tag present in ChiZ-FL (but absent in ChiZ1-64) might attach to the membrane. 21 That still leaves two RP residues, Arg34 and Arg39, in the midsection unaccounted for. As the data from the next experiment indicate, even in the ChiZ1-64 construct, these two residues, along with other midsection Arg residues, interact with lipids, and the resulting loss in dynamics potentially prevented their detection by INEPT, but the loss in dynamics was incomplete so Arg34 and Arg39 were not detectable by CP either.…”

Section: Resultsmentioning

confidence: 99%

Fuzzy Association of an Intrinsically Disordered Protein with Acidic Membranes

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Cross

et al. 2020

JACS Au

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Many physiological and pathophysiological processes, including Mycobacterium tuberculosis ( Mtb ) cell division, may involve fuzzy membrane association by proteins via intrinsically disordered regions. The fuzziness is extreme when the conformation and pose of the bound protein and the composition of the proximal lipids are all highly dynamic. Here, we tackled the challenge in characterizing the extreme fuzzy membrane association of the disordered, cytoplasmic N-terminal region (NT) of ChiZ, an Mtb divisome protein, by combining solution and solid-state NMR spectroscopy and molecular dynamics simulations. While membrane-associated NT does not gain any secondary structure, its interactions with lipids are not random, but formed largely by Arg residues predominantly in the second, conserved half of the NT sequence. As NT frolics on the membrane, lipids quickly redistribute, with acidic lipids, relative to zwitterionic lipids, preferentially taking up Arg-proximal positions. The asymmetric engagement of NT arises partly from competition between acidic lipids and acidic residues, all in the first half of NT, for Arg interactions. This asymmetry is accentuated by membrane insertion of the downstream transmembrane helix. This type of semispecific molecular recognition may be a general mechanism by which disordered proteins target membranes.

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Section: Resultsmentioning

confidence: 99%

Fuzzy Association of an Intrinsically Disordered Protein with Acidic Membranes

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Cross

et al. 2020

JACS Au

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“…The amphipathic α-helical secondary structure presented by several AMPs is crucial for peptide activity. Some reports already explored piscidin conformation and activity, as well as their mode of action on pathogen membranes, using artificial membranes that mimic the natural ones and can elucidate the mechanisms underlying the effects observed on pathogenic cells [47][48][49]. In addition, Schiffer-Edmundson helical wheel diagrams are often used to predict the hydrophobic and hydrophilic regions in the secondary structure of these peptides.…”

Section: Discussionmentioning

confidence: 99%

The Diverse Piscidin Repertoire of the European Sea Bass (Dicentrarchus labrax): Molecular Characterization and Antimicrobial Activities

Barroso

Carvalho

et al. 2020

IJMS

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Fish rely on their innate immune responses to cope with the challenging aquatic environment, with antimicrobial peptides (AMPs) being one of the first line of defenses. Piscidins are a group of fish specific AMPs isolated in several species. However, in the European sea bass (Dicentrarchus labrax), the piscidin family remains poorly understood. We identified six different piscidins in sea bass, performed an in-depth molecular characterization and evaluated their antimicrobial activities against several bacterial and parasitic pathogens. Sea bass piscidins present variable amino acid sequences and antimicrobial activities, and can be divided in different sub groups: group 1, formed by piscidins 1 and 4; group 2, constituted by piscidins 2 and 5, and group 3, formed by piscidins 6 and 7. Additionally, we demonstrate that piscidins 1 to 5 possess a broad effect on multiple microorganisms, including mammalian parasites, while piscidins 6 and 7 have poor antibacterial and antiparasitic activities. These results raise questions on the functions of these peptides, particularly piscidins 6 and 7. Considering their limited antimicrobial activity, these piscidins might have other functional roles, but further studies are necessary to better understand what roles might those be.

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“…We also observed that after exposure to 500 nM NEC220, which deposited NEC220 on the membrane surface at high density (molar ratio protein/lipid (P/L) = 1/45), the membrane thickened by 0.49 ± 0.17 Å (68% confidence interval), in the context of the orientation model (Table S1). Thinning of membranes tethered to flat substrates has been observed with proteins that generate positive curvature by inserting into the headgroup region (Chen et al, 2003;Mihailescu et al, 2014;Mihailescu et al, 2019). This is because forcing the headgroups apart on a flat substrate increases the area per lipid and thins the membrane (the hydrophobic tails form a constant-volume cylinder, the height of which must decrease if the area is increased).…”

Section: Soluble Nec Inserts Peripherally Into the Tethered Lipid Bilayersmentioning

confidence: 99%

Highly basic clusters in the HSV-1 nuclear egress complex drive membrane budding by inducing lipid ordering

Lai

et al. 2021

Preprint

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During replication of herpesviruses, capsids escape from the nucleus into the cytoplasm by budding at the inner nuclear membrane. This unusual process is mediated by the viral nuclear egress complex (NEC) that deforms the membrane around the capsid by oligomerizing into a hexagonal, membrane-bound scaffold. Here, we found that highly basic membrane-proximal regions (MPRs) of the NEC alter lipid order by inserting into the lipid headgroups and also promote negative Gaussian curvature. We also find that the electrostatic interactions between the MPRs and the membranes are essential for membrane deformation. One of the MPRs is phosphorylated by a viral kinase during infection, and the corresponding phosphomimicking mutations block capsid nuclear egress. We show that the same phosphomimicking mutations disrupt the NEC/membrane interactions and inhibit NEC-mediated budding in vitro, providing a biophysical explanation for the in-vivo phenomenon. Our data suggest that the NEC generates negative membrane curvature by both lipid ordering and protein scaffolding and that phosphorylation acts as an "off" switch that inhibits the membrane-budding activity of the NEC to prevent capsid-less budding.

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

Cited by 64 publications

References 122 publications

Fuzzy Association of an Intrinsically Disordered Protein with Acidic Membranes

Fuzzy Association of an Intrinsically Disordered Protein with Acidic Membranes

The Diverse Piscidin Repertoire of the European Sea Bass (Dicentrarchus labrax): Molecular Characterization and Antimicrobial Activities

Highly basic clusters in the HSV-1 nuclear egress complex drive membrane budding by inducing lipid ordering

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