Artificially Induced Protein–Membrane Anchorage with Cholesterol‐Based Recognition Agents as a New Therapeutic Concept

Avadisian, Miriam; Fletcher, Steven; Liu, Baoxu; Zhao, Wei; Yue, Peibin; Badali, Daniel; Xu, Wei; Schimmer, Aaron D.; Turkson, James; Gradinaru, Claudiu C.; Gunning, Patrick T.

doi:10.1002/anie.201102486

Cited by 19 publications

(20 citation statements)

References 22 publications

(14 reference statements)

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“…A single anchoring moiety is sufficient for proteins considerably larger in size [50] than even large copy number multimeric inhibitors with a monomer size of 35–40 amino acids. Data published after our work was completed lend further support to our hypothesis [51]. Moreover, we envisage having only two copies of the inhibitor sequence, as discussed below.…”

Section: Resultssupporting

confidence: 73%

A General Strategy to Endow Natural Fusion-protein-Derived Peptides with Potent Antiviral Activity

Pessi

Langella

Capitò³

et al. 2012

PLoS ONE

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Fusion between the viral and target cell membranes is an obligatory step for the infectivity of all enveloped virus, and blocking this process is a clinically validated therapeutic strategy.Viral fusion is driven by specialized proteins which, although specific to each virus, act through a common mechanism, the formation of a complex between two heptad repeat (HR) regions. The HR regions are initially separated in an intermediate termed “prehairpin”, which bridges the viral and cell membranes, and then fold onto each other to form a 6-helical bundle (6HB), driving the two membranes to fuse. HR-derived peptides can inhibit viral infectivity by binding to the prehairpin intermediate and preventing its transition to the 6HB.The antiviral activity of HR-derived peptides differs considerably among enveloped viruses. For weak inhibitors, potency can be increased by peptide engineering strategies, but sequence-specific optimization is time-consuming. In seeking ways to increase potency without changing the native sequence, we previously reported that attachment to the HR peptide of a cholesterol group (”cholesterol-tagging”) dramatically increases its antiviral potency, and simultaneously increases its half-life in vivo. We show here that antiviral potency may be increased by combining cholesterol-tagging with dimerization of the HR-derived sequence, using as examples human parainfluenza virus, Nipah virus, and HIV-1. Together, cholesterol-tagging and dimerization may represent strategies to boost HR peptide potency to levels that in some cases may be compatible with in vivo use, possibly contributing to emergency responses to outbreaks of existing or novel viruses.

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Section: Resultssupporting

confidence: 73%

A General Strategy to Endow Natural Fusion-protein-Derived Peptides with Potent Antiviral Activity

Pessi

Langella

Capitò³

et al. 2012

PLoS ONE

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“…After incubation for about 2 h at room temperature, the coverslips were separated and rinsed with PBS buffer thoroughly [30]. The formation of supported lipid bilayer (SLB) on the glass coverslip was verified by fluorescence recovery after photobleaching (FRAP) experiments using a mixture of 90 mol% DOPC and 10 mol% Bodipy-FL lipids (Movie S1, Supporting Information (SI)).…”

Section: Sample Preparationmentioning

confidence: 99%

“…Then the coverslip was rinsed with PBS buffer to remove the non-attached dye molecules. The SLB was then imaged on a TIRF microscope [30] and single fluorophores that remained immobilized on the SLB were identified and counted in a region of interest of 36 × 36 μm 2 . At least three different samples and five distinct regions in each sample were recorded and analyzed.…”

Section: Single-molecule Imaging Of Fluorophores On Slbsmentioning

confidence: 99%

Choosing the right fluorophore for single-molecule fluorescence studies in a lipid environment

Zhang

Yomo

Gradinaru

2017

Biochimica et Biophysica Acta (BBA) - Biomembranes

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Nonspecific interactions between lipids and fluorophores can alter the outcomes of single-molecule spectroscopy of membrane proteins in live cells, liposomes or lipid nanodiscs and of cytosolic proteins encapsulated in liposomes or tethered to supported lipid bilayers. To gain insight into these effects, we examined interactions between 9 dyes that are commonly used as labels for single-molecule fluorescence (SMF) and 6 standard lipids including cationic, zwitterionic and anionic types. The diffusion coefficients of dyes in the absence and presence of set amounts of lipid vesicles were measured by fluorescence correlation spectroscopy (FCS). The partition coefficients and the free energies of partitioning for different fluorophore-lipid pairs were obtained by global fitting of the titration FCS curves. Lipids with different charges, head groups and degrees of chain saturation were investigated, and interactions with dyes are discussed in terms of hydrophobic, electrostatic and steric contributions. Fluorescence imaging of individual fluorophores adsorbed on supported lipid bilayers provides visualization and additional quantification of the strength of dye-lipid interaction in the context of single-molecule measurements. By dissecting fluorophore-lipid interactions, our study provides new insights into setting up single-molecule fluorescence spectroscopy experiments with minimal interference from interactions between fluorescent labels and lipids in the environment.

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“…Dye 3 , a more lipophilic version of 1 , was synthesized by a similar procedure, and then converted into the cholesterol conjugate 4 by amide bond coupling with an amine functionalized cholesterol derivative. 21 …”

Section: Resultsmentioning

confidence: 99%

Using membrane composition to fine-tune the pK_a of an optical liposome pH sensor

et al. 2016

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Liposomes containing membrane-anchored pH-sensitive optical probes are valuable sensors for monitoring pH in various biomedical samples. The dynamic range of the sensor is maximized when the probe pKa is close to the expected sample pH. While some biomedical samples are close to neutral pH there are several circumstances where the pH is 1 or 2 units lower. Thus, there is a need to fine-tune the probe pKa in a predictable way. This investigation examined two lipid-conjugated optical probes, each with appended deep-red cyanine dyes containing indoline nitrogen atoms that are protonated in acid. The presence of anionic phospholipids in the liposomes stabilized the protonated probes and increased the probe pKa values by < 1 unit. The results show that rational modification of the membrane composition is a general non-covalent way to fine-tune the pKa of an optical liposome sensor for optimal pH sensing performance.

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Artificially Induced Protein–Membrane Anchorage with Cholesterol‐Based Recognition Agents as a New Therapeutic Concept

Cited by 19 publications

References 22 publications

A General Strategy to Endow Natural Fusion-protein-Derived Peptides with Potent Antiviral Activity

A General Strategy to Endow Natural Fusion-protein-Derived Peptides with Potent Antiviral Activity

Choosing the right fluorophore for single-molecule fluorescence studies in a lipid environment

Using membrane composition to fine-tune the pK_a of an optical liposome pH sensor

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Artificially Induced Protein–Membrane Anchorage with Cholesterol‐Based Recognition Agents as a New Therapeutic Concept

Cited by 19 publications

References 22 publications

A General Strategy to Endow Natural Fusion-protein-Derived Peptides with Potent Antiviral Activity

A General Strategy to Endow Natural Fusion-protein-Derived Peptides with Potent Antiviral Activity

Choosing the right fluorophore for single-molecule fluorescence studies in a lipid environment

Using membrane composition to fine-tune the pKa of an optical liposome pH sensor

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Product

Resources

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Using membrane composition to fine-tune the pK_a of an optical liposome pH sensor