Liposome fusion with orthogonal coiled coil peptides as fusogens: the efficacy of roleplaying peptides

Daudey, Geert A.; Shen, Ming; Singhal, Ankush; Est, Patrick van der; Sevink, G. J. A.; Boyle, Aimee L.; Kros, Alexander

doi:10.1039/d0sc06635d

Cited by 17 publications

(27 citation statements)

References 76 publications

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“…Thus, E/K peptides have the potential to be employed for drug delivery in biological systems as evidenced by the efficient targeted delivery of the small molecule anionionophore into GUVs loaded into LUVs [ 127 ]. The E/K system can be further extended by the development of a small library of lipidated coiled-coil-forming orthogonal peptides, with implications for membrane fusion specificity [ 128 ].…”

Section: Snare-mimetics Functional Peptides Mediating Membrane Fusionmentioning

confidence: 99%

SNARE Modulators and SNARE Mimetic Peptides

Khvotchev

Soloviev

2022

Biomolecules

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The soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein (SNAP) receptor (SNARE) proteins play a central role in most forms of intracellular membrane trafficking, a key process that allows for membrane and biocargo shuffling between multiple compartments within the cell and extracellular environment. The structural organization of SNARE proteins is relatively simple, with several intrinsically disordered and folded elements (e.g., SNARE motif, N-terminal domain, transmembrane region) that interact with other SNAREs, SNARE-regulating proteins and biological membranes. In this review, we discuss recent advances in the development of functional peptides that can modify SNARE-binding interfaces and modulate SNARE function. The ability of the relatively short SNARE motif to assemble spontaneously into stable coiled coil tetrahelical bundles has inspired the development of reduced SNARE-mimetic systems that use peptides for biological membrane fusion and for making large supramolecular protein complexes. We evaluate two such systems, based on peptide-nucleic acids (PNAs) and coiled coil peptides. We also review how the self-assembly of SNARE motifs can be exploited to drive on-demand assembly of complex re-engineered polypeptides.

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Section: Snare-mimetics Functional Peptides Mediating Membrane Fusionmentioning

confidence: 99%

SNARE Modulators and SNARE Mimetic Peptides

Khvotchev

Soloviev

2022

Biomolecules

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“…[30][31][32] Peptide K is an amphipathic helical peptide and was specifically designed to interact with peptide E, but when confined to a membrane, it also interacts with lipid bilayers. [33][34][35][36][37] Due to the dual affinity of peptide K to both peptide E and lipid membranes, we herein investigate whether dimerization of peptide K could enhance liposomal drug delivery to cells. The influence of peptide dimerization on solution properties was studied, in addition to the ability to induce fusion of liposomes with cells to facilitate drug delivery (Scheme 1).…”

Section: Introductionmentioning

confidence: 99%

Enhanced liposomal drug delivery via membrane fusion triggered by dimeric coiled-coil peptides

Zeng

Shen

Singhal

et al. 2023

Preprint

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An ideal nanomedicine system improves the therapeutic efficacy of a drug. However, most nanomedicines enter the cell via endosomal/lysosomal pathways and typically only a small fraction of the cargo enters the cytosol inducing a therapeutic effect. To circumvent these inefficient drug delivery pathways, alternative approaches are desired. Inspired by fusion machinery found in nature, various synthetic fusogens have been developed to mediate the fusion of vesicles. Previously we used lipidated versions of the heterodimeric peptide pair E4/K4 to induce membrane fusion and applied this approach to deliver drugs in vitro and in vivo. Peptide K4 was designed to interact specifically with peptide E, but it was also shown to have an affinity for lipid membranes resulting in membrane remodeling. As the next step towards efficient in vivo membrane fusion for potential therapeutic applications, our rationale was to design efficient fusogen variants that could have multiple interactions simultaneously. To achieve this goal, we synthesized dimeric coiled-coil peptide variants of peptide K to facilitate improved fusion with peptide E-modified liposomes and mammalian cells. The secondary structure and self-assembly of various dimers were studied; the parallel PK4 dimer forms temperature-dependent higher-order assemblies, while linear K4 dimers form tetramer-like homodimers. The findings regarding the structures and membrane interactions of the PK4 dimer were supported by molecular dynamics simulations. Upon addition of the complementary peptide E4, the parallel PK4 dimer induced the strongest coiled-coil interaction resulting in a higher cellular uptake of the liposome-encapsulated cargo, as compared to the linear dimer and monomeric designs. Using a wide spectrum of endocytosis inhibitors, it was shown that membrane fusion was the main cellular uptake pathway. Delivery of the antitumor drug doxorubicin (DOX) resulted in efficient cellular delivery and concomitant antitumor efficacy. These findings not only offer important mechanistic insights into the design of coiled-coil driven membrane fusion systems but also provide novel strategies for developing peptide-based biomaterials.

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“…Expansion of the fusion stalk results in a hemifusion diaphragm (HD) until formation of a fusion pore in the HD completes the fusion reaction. Many details of the underlying molecular mechanisms have been addressed using simpler lipidated model systems including small molecules, coiled-coil structures, − or complementary DNA − or peptide nucleic acid strands …”

Section: Introductionmentioning

confidence: 99%

“…We demonstrate that such novel biotin-modified AB 5 cholera toxin–streptavidin complexes [Strep–(AB 5 ) n ] exhibit both cross-linking and fusogenic functions. In contrast to the aforementioned fusogenic strategies in which complementary recognition elements are lipidated and introduced into separate vesicles, − the Strep–(AB 5 ) n is a soluble protein; that is, it is neither embedded nor covalently attached to the membrane but binds non-covalently to liposomes containing the GM1 ganglioside. Streptavidin-mediated cross-linking of two or more AB 5 complexes allows the assemblies to bind to two opposing membranes in parallel, which is the first prerequisite for fusion.…”

Section: Introductionmentioning

confidence: 99%

Membrane Fusion Mediated by Non-covalent Binding of Re-engineered Cholera Toxin Assemblies to Glycolipids

et al. 2022

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Membrane fusion is essential for the transport of macromolecules and viruses across membranes. While glycan-binding proteins (lectins) often initiate cellular adhesion, subsequent fusion events require additional protein machinery. No mechanism for membrane fusion arising from simply a protein binding to membrane glycolipids has been described thus far. Herein, we report that a biotinylated protein derived from cholera toxin becomes a fusogenic lectin upon cross-linking with streptavidin. This novel reengineered protein brings about hemifusion and fusion of vesicles as demonstrated by mixing of fluorescently labeled lipids between vesicles as well as content mixing of liposomes filled with fluorescently labeled dextran. Exclusion of the complex at vesicle−vesicle interfaces could also be observed, indicating the formation of hemifusion diaphragms. Discovery of this fusogenic lectin complex demonstrates that new emergent properties can arise from simple changes in protein architecture and provides insights into new mechanisms of lipid-driven fusion.

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Liposome fusion with orthogonal coiled coil peptides as fusogens: the efficacy of roleplaying peptides

Abstract: Biological membrane fusion is a highly specific and coordinated process as a multitude of vesicular fusion events proceed simultaneously in a complex environment with minimal off-target delivery. In this study,...

Cited by 17 publications

References 76 publications

SNARE Modulators and SNARE Mimetic Peptides

SNARE Modulators and SNARE Mimetic Peptides

Enhanced liposomal drug delivery via membrane fusion triggered by dimeric coiled-coil peptides

Membrane Fusion Mediated by Non-covalent Binding of Re-engineered Cholera Toxin Assemblies to Glycolipids

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