A non-zipper-like tetrameric coiled coil promotes membrane fusion

Zheng, Tingting; Bulacu, Monica; Daudey, Geert A.; Versluis, Frank; Voskuhl, Jens; Martelli, Giuliana; Raap, J.; Sevink, G. J. A.; Kros, Alexander; Boyle, Aimee L.

doi:10.1039/c5ra26175a

Cited by 21 publications

(20 citation statements)

References 53 publications

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“…The formation of a heterodimeric coiled-coil brings the two opposing membranes into close proximity, inducing efficient, leakage-free, membrane fusion 25 . Numerous aspects of this system, including: the length and oligomer state of the peptides 26,27 , the PEG spacer length 28 , the nature and position of the lipid anchor [29][30][31] , and the effects of these variables on the fusion process have been investigated. Moreover, the system has been employed for the targeted delivery of various cargoes to the membranes of GUVs and cells [32][33][34][35] .…”

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confidence: 99%

Controlled Peptide-Mediated Vesicle Fusion Assessed by Simultaneous Dual-Colour Time-Lapsed Fluorescence Microscopy

Mora

Boyle

Kolck

et al. 2020

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We have employed a model system, inspired by SnARe proteins, to facilitate membrane fusion between Giant Unilamellar Vesicles (GUVs) and Large Unilamellar Vesicles (LUVs) under physiological conditions. in this system, two synthetic lipopeptide constructs comprising the coiled-coil heterodimerforming peptides K 4 , (KiAALKe) 4 , or e 4 , (eiAALeK) 4 , a peG spacer of variable length, and a cholesterol moiety to anchor the peptides into the liposome membrane replace the natural SnARe proteins. GUVs are functionalized with one of the lipopeptide constructs and the fusion process is triggered by adding LUVs bearing the complementary lipopeptide. Dual-colour time lapse fluorescence microscopy was used to visualize lipid-and content-mixing. Using conventional confocal microscopy, lipid mixing was observed on the lipid bilayer of individual GUVs. in addition to lipid-mixing, content-mixing assays showed a low efficiency due to clustering of K 4-functionalized LUVs on the GUVs target membranes. We showed that, through the use of the non-ionic surfactant Tween 20, content-mixing between GUVs and LUVs could be improved, meaning this system has the potential to be employed for drug delivery in biological systems.

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confidence: 99%

Controlled Peptide-Mediated Vesicle Fusion Assessed by Simultaneous Dual-Colour Time-Lapsed Fluorescence Microscopy

Mora

Boyle

Kolck

et al. 2020

Sci Rep

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“…Parameters foe the DLS measurements were consistent with previous publications. [21][22][23][24][25] A 633 nm laser and 173 scattering angle were set as xed parameters. For each measurement, the sample was allowed to have 2 min of equilibration time.…”

Section: Cell Culturementioning

confidence: 99%

Berberine nanoparticles for promising sonodynamic therapy of a HeLa xenograft tumour

et al. 2019

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“…Inspired by this natural and highly controlled transport mechanism, supramolecular and biomaterials chemists designed synthetic targeted membrane fusion systems in order to study the mechanism of membrane fusion at a fundamental level, aiming to explore applications in the design nanocarriers for drug delivery systems. Our previously published synthetic membrane fusion system shows an effective way of inducing liposome-liposome fusion [10,[14][15][16][17][18][19][20][21]. There are three key components in the design of our fusion model, including molecular recognition composed of two complimentary coiled coil forming peptides (E/K), a flexible linker (PEG) and a lipid (L) anchor (DOPE).…”

Section: Introductionmentioning

confidence: 99%

High efficiency liposome fusion induced by reducing undesired membrane peptides interaction

et al. 2019

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A full membrane fusion model which attains both complete lipid mixing and content mixing liposomal membranes mediated by coiled-coil forming lipopeptides LPK [L-PEG12-(KIAALKE)3] and LPE [L-PEG12-(EIAALEK)3] is presented. The electrostatic effects of lipid anchored peptides on fusion efficiency was investigated. For this, the original amino acid sequence of the membrane bound LPK was varied at its ‘f’-position of the helical structure, i.e. via mutating the anionic glutamate residues by either neutral serines or cationic lysines. Both CD and fluorescence measurements showed that replacing the negatively charged glutamate did not significantly alter the peptide ability to form a coiled coil, but lipid mixing and content mixing assays showed more efficient liposome-liposome fusion resulting in almost quantitative content mixing for the lysine mutated analogue (LPKK) in conjunction with LPE. A mechanism is proposed for a fusion model triggered by membrane destabilizing effects mediated by the membrane destabilizing activety of LPK in cooperation with the electrostatic activity of LPE. This new insight may enlightens the further development of a promising nano carrier tool for biomedical applications.

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A non-zipper-like tetrameric coiled coil promotes membrane fusion

Abstract: A parallel heterodimeric coiled coil can be mutated to an antiparallel tetrameric species by reversing the sequences of one of the peptides. This tetramer is capable of facilitating fast, efficient, membrane fusion of liposomes.

Cited by 21 publications

References 53 publications

Controlled Peptide-Mediated Vesicle Fusion Assessed by Simultaneous Dual-Colour Time-Lapsed Fluorescence Microscopy

Controlled Peptide-Mediated Vesicle Fusion Assessed by Simultaneous Dual-Colour Time-Lapsed Fluorescence Microscopy

Berberine nanoparticles for promising sonodynamic therapy of a HeLa xenograft tumour

High efficiency liposome fusion induced by reducing undesired membrane peptides interaction

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