In Vitro and In Vivo Supramolecular Modification of Biomembranes Using a Lipidated Coiled‐Coil Motif

Zope, Harshal; Versluis, Frank; Ordas, Anita; Voskuhl, Jens; Spaink, Herman P.; Kros, Alexander

doi:10.1002/anie.201306033

Cited by 57 publications

(125 citation statements)

References 48 publications

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“…Fig. 2 A displays the change of fluorescence intensity upon varying the lipid to peptide ratio by addition of vesicles of the composition DOPC:DOPE:cholesterol (molar ratio: 2:1:1), as used in recent vesicle fusion studies (5)(6)(7)(8)(9)(10)16,44,45). For comparative purposes, the total concentration of the tryptophan bearing peptide (X*, X ¼ K or E, respectively) was kept constant.…”

Section: Membrane Binding Of K Before and After Vesicle Dockingmentioning

confidence: 99%

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A Coiled-Coil Peptide Shaping Lipid Bilayers upon Fusion

Rabe

Aisenbrey

Pluháčková

et al. 2016

Biophysical Journal

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A system based on two designed peptides, namely the cationic peptide K, (KIAALKE) 3 , and its complementary anionic counterpart called peptide E, (EIAALEK) 3 , has been used as a minimal model for membrane fusion, inspired by SNARE proteins. Although the fact that docking of separate vesicle populations via the formation of a dimeric E/K coiled-coil complex can be rationalized, the reasons for the peptides promoting fusion of vesicles cannot be fully explained. Therefore it is of significant interest to determine how the peptides aid in overcoming energetic barriers during lipid rearrangements leading to fusion. In this study, investigations of the peptides' interactions with neutral PC/PE/cholesterol membranes by fluorescence spectroscopy show that tryptophan-labeled K* binds to the membrane (K K*~6 .2 10 3 M À1 ), whereas E* remains fully water-solvated. 15 N-NMR spectroscopy, depth-dependent fluorescence quenching, CD-spectroscopy experiments, and MD simulations indicate a helix orientation of K* parallel to the membrane surface. Solid-state 31 P-NMR of oriented lipid membranes was used to study the impact of peptide incorporation on lipid headgroup alignment. The membrane-immersed K* is found to locally alter the bilayer curvature, accompanied by a change of headgroup orientation relative to the membrane normal and of the lipid composition in the vicinity of the bound peptide. The NMR results were supported by molecular dynamics simulations, which showed that K reorganizes the membrane composition in its vicinity, induces positive membrane curvature, and enhances the lipid tail protrusion probability. These effects are known to be fusion relevant. The combined results support the hypothesis for a twofold role of K in the mechanism of membrane fusion: 1) to bring opposing membranes into close proximity via coiled-coil formation and 2) to destabilize both membranes thereby promoting fusion.

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Section: Membrane Binding Of K Before and After Vesicle Dockingmentioning

confidence: 99%

“…Therefore for further quenching measurements at constant lipid concentrations, this ratio was chosen. For measurements with LPE* and LPK*, a ratio of 100:1 was chosen because this reflects the conditions of typical vesicle fusion experiments (5)(6)(7)(8)(9)(10)16,44,45).…”

Section: Membrane Binding Of K Before and After Vesicle Dockingmentioning

confidence: 99%

A Coiled-Coil Peptide Shaping Lipid Bilayers upon Fusion

Rabe

Aisenbrey

Pluháčková

et al. 2016

Biophysical Journal

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“…The lipopeptides were analyzed by electrospray-ionization mass spectrometry (E3Cys-MCCDOPE: 3882 m z À1 [Mþ], K3Cys-MCCDOPE: 3827 m z À1 [Mþ]). Additionally, cholesterol-PEG12-E4 and cholesterol-PEG12-K4 were used to induce membrane fusion between the silica beads (10)(11)(12)(13). some experiments one fraction contained a different fluorescent dye lipid such as Oregon Green 488 DPPE, Atto488-DOPE, or ATTO390DOPE (1 mol %).…”

Section: Lipopeptidesmentioning

confidence: 99%

Geometry of the Contact Zone between Fused Membrane-Coated Beads Mimicking Cell-Cell Fusion

Savić

Kliesch

Verbeek

et al. 2016

Biophysical Journal

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The fusion of lipid membranes is a key process in biology. It enables cells and organelles to exchange molecules with their surroundings, which otherwise could not cross the membrane barrier. To study such complex processes we use simplified artificial model systems, i.e., an optical fusion assay based on membrane-coated glass spheres. We present a technique to analyze membrane-membrane interactions in a large ensemble of particles. Detailed information on the geometry of the fusion stalk of fully fused membranes is obtained by studying the diffusional lipid dynamics with fluorescence recovery after photobleaching experiments. A small contact zone is a strong obstruction for the particle exchange across the fusion spot. With the aid of computer simulations, fluorescence-recovery-after-photobleaching recovery times of both fused and single-membrane-coated beads allow us to estimate the size of the contact zones between two membrane-coated beads. Minimizing delamination and bending energy leads to minimal angles close to those geometrically allowed.

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“…7 Very recently Kros and co-workers reported a liposome fusion process to introduce cholesterol functionalized coiled-coil forming peptides, which allowed for the in vivo decoration of cellular membranes. 8 In prior work by Yousaf and co-workers, such liposome fusion strategies were shown to yield three-dimensional tissue-like structures. 9 These studies not only show us that innovative approaches are required to address cells at the molecular level but also stress that we need to clearly understand the nature of the interactions B between our synthetic molecules and complex cellular environments.…”

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confidence: 97%

Incorporating Bacteria as a Living Component in Supramolecular Self-Assembled Monolayers through Dynamic Nanoscale Interactions

2015

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Supramolecular assemblies, formed through noncovalent interactions, has become particularly attractive to develop dynamic and responsive architectures to address living systems at the nanoscale. Cucurbit[8]uril (CB[8]), a pumpkin shaped macrocylic host molecule, has been successfully used to construct various self-assembled architectures for biomedical applications since it can simultaneously bind two aromatic guest molecules within its cavity. Such architectures can also be designed to respond to external stimuli. Integrating living organisms as an active component into such supramolecular architectures would add a new dimension to the capabilities of such systems. To achieve this, we have incorporated supramolecular functionality at the bacterial surface by genetically modifying a transmembrane protein to display a CB[8]-binding motif as part of a cystine-stabilized miniprotein. We were able to confirm that this supramolecular motif on the bacterial surface specifically binds CB[8] and forms multiple intercellular ternary complexes leading to aggregation of the bacterial solution. We performed various aggregation experiments to understand how CB[8] interacts with this bacterial strain and also demonstrate that it can be chemically reversed using a competitor. To confirm that this strain can be incorporated with a CB[8] based architecture, we show that the bacterial cells were able to adhere to CB[8] self-assembled monolayers (SAMs) on gold and still retain considerable motility for several hours, indicating that the system can potentially be used to develop supramolecular bacterial biomotors. The bacterial strain also has the potential to be combined with other CB[8] based architectures like nanoparticles, vesicles and hydrogels.

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In Vitro and In Vivo Supramolecular Modification of Biomembranes Using a Lipidated Coiled‐Coil Motif

Cited by 57 publications

References 48 publications

A Coiled-Coil Peptide Shaping Lipid Bilayers upon Fusion

A Coiled-Coil Peptide Shaping Lipid Bilayers upon Fusion

Geometry of the Contact Zone between Fused Membrane-Coated Beads Mimicking Cell-Cell Fusion

Incorporating Bacteria as a Living Component in Supramolecular Self-Assembled Monolayers through Dynamic Nanoscale Interactions

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