Transmembrane Domain Peptide/Peptide Nucleic Acid Hybrid as a Model of a SNARE Protein in Vesicle Fusion

Lygina, Antonina S.; Meyenberg, Karsten; Jahn, Reinhard; Diederichsen, Ulf

doi:10.1002/anie.201101951

Cited by 57 publications

(76 citation statements)

References 29 publications

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“…Presently we do not know yet in which way the proteins interact which each other, which of the coiled-coils are participating, and to what extent the hybrid complexes are structurally similar to the highly conserved SNARE four-helix bundles. However, it appears that the structural tolerance of the SNARE-based zipper mechanism for membrane fusion is much higher than anticipated (45), which is also supported by the surprising structural diversity of artificial SNARE mimetics capable of inducing fusion in vitro by some kind of zippering mechanism (46,47). Further structural investigation of the LegC/SNARE hybrid complex can thus be expected to yield novel insights not only into the degree of structural conservation but also into the mechanisms involved in SNARE-mediated membrane fusion.…”

Section: Discussionmentioning

confidence: 92%

Direct targeting of membrane fusion by SNARE mimicry: Convergent evolution ofLegionellaeffectors

Shi

Halder

Yavuz

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Legionella pneumophila, the Gram-negative pathogen causing Legionnaires’ disease, infects host cells by hijacking endocytic pathways and forming a Legionella-containing vacuole (LCV) in which the bacteria replicate. To promote LCV expansion and prevent lysosomal targeting, effector proteins are translocated into the host cell where they alter membrane traffic. Here we show that three of these effectors [LegC2 (Legionella eukaryotic-like gene C2)/YlfB (yeast lethal factor B), LegC3, and LegC7/YlfA] functionally mimic glutamine (Q)-SNARE proteins. In infected cells, the three proteins selectively form complexes with the endosomal arginine (R)-SNARE vesicle-associated membrane protein 4 (VAMP4). When reconstituted in proteoliposomes, these proteins avidly fuse with liposomes containing VAMP4, resulting in a stable complex with properties resembling canonical SNARE complexes. Intriguingly, however, the LegC/SNARE hybrid complex cannot be disassembled by N-ethylmaleimide-sensitive factor. We conclude that LegCs use SNARE mimicry to divert VAMP4-containing vesicles for fusion with the LCV, thus promoting its expansion. In addition, the LegC/VAMP4 complex avoids the host’s disassembly machinery, thus effectively trapping VAMP4 in an inactive state.

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Section: Discussionmentioning

confidence: 92%

Direct targeting of membrane fusion by SNARE mimicry: Convergent evolution ofLegionellaeffectors

Shi

Halder

Yavuz

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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“…[11]. The sequences of the transmembrane domains (TMDs) of the model peptides were from Syntaxin-1A (256-288) and Synaptobrevin-2 (96-116) from rattus norvegicus.…”

Section: Synthetic Proceduresmentioning

confidence: 99%

“…[11] was followed with the following adaptations being made: The stock solutions of the reagents were prepared freshly each day. The resin which contained the transmembrane domain (1.0 eq) was swollen 2 h in NMP before the SPPS cycle started.…”

Section: Synthetic Proceduresmentioning

confidence: 99%

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PNA‐Hybridsequenzen als Erkennungseinheiten in SNARE‐Protein‐analogen Peptiden

et al. 2018

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Die Fusion von Membranen ist ein grundlegender Prozess in der Natur, der oft durch die spezifische Interaktion von SNARE‐Proteinen herbeigeführt wird. SNARE‐Modellsysteme, in denen die SNARE‐Domänen durch kleine artifizielle Einheiten ersetzt sind, stellen wertvolle Werkzeuge dar, um die Membranfusion in vitro zu untersuchen. Hier werden Synthese und Analyse von SNARE‐Modellpeptiden vorgestellt, die eine aus zwei unterschiedlichen Peptidnukleinsäure(PNA)‐Topologien zusammengesetzte Erkennungseinheit besitzen. Diese neuartige Erkennungseinheit wurde entwickelt, um den Reißverschluss‐Mechanismus der SNARE‐Proteine nachzustellen, der für die Einleitung der SNARE‐Protein vermittelten Membranfusion verantwortlich gemacht wird. Die Erkennungseinheit besteht aus N‐(2‐Aminoethyl)glycin‐PNA (Aeg‐PNA) und Alanyl‐PNA, die beide ihren entsprechend komplementären Strang erkennen können, allerdings unterschiedliche Duplextopologien und Duplexbildungskinetiken aufweisen. Neben der Charakterisierung der Duplexbildung der PNA‐Hybridoligomere wird auch die Fusogenität der Modellpeptide anhand von Lipidmischungsexperimenten untersucht und es wird gezeigt, dass die Modellpeptide Fusion induzieren. Unerwartet war, dass Peptide mit einer Erkennungseinheit von nur fünf Aeg‐PNA‐Einheiten bereits die Fusion von Liposomen induzieren und sich dabei bislang sogar als am effizientesten erwiesen haben.

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“…[11,12] By combining proteins with synthetic supramolecular systems,w estand to benefit from the unique structural and functional features of both, and broaden the functionality of signaling systems.Notwithstanding the level of sophistication and control in natural systems, [13,14] impressive first examples of supramolecular systems on the way to these goals have been reported. [15][16][17] Cucurbit [8]uril (Q8) is ac yclic glycoluril-derived supramolecular host system capable of binding simultaneously to two N-terminal phenylalanine residues. [18] Q8 has demonstrated significant potential as ascaffold for the formation of supramolecular protein complexes, [19,20] as well as to modulate the function of biomaterials.…”

Section: Supramolecular Control Over Split-luciferase Complementationmentioning

confidence: 99%

Supramolecular Control over Split‐Luciferase Complementation

et al. 2016

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Supramolecular split‐enzyme complementation restores enzymatic activity and allows for on–off switching. Split‐luciferase fragment pairs were provided with an N‐terminal FGG sequence and screened for complementation through host‐guest binding to cucurbit[8]uril (Q8). Split‐luciferase heterocomplex formation was induced in a Q8 concentration dependent manner, resulting in a 20‐fold upregulation of luciferase activity. Supramolecular split‐luciferase complementation was fully reversible, as revealed by using two types of Q8 inhibitors. Competition studies with the weak‐binding FGG peptide revealed a 300‐fold enhanced stability for the formation of the ternary heterocomplex compared to binding of two of the same fragments to Q8. Stochiometric binding by the potent inhibitor memantine could be used for repeated cycling of luciferase activation and deactivation in conjunction with Q8, providing a versatile module for in vitro supramolecular signaling networks.

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Transmembrane Domain Peptide/Peptide Nucleic Acid Hybrid as a Model of a SNARE Protein in Vesicle Fusion

Cited by 57 publications

References 29 publications

Direct targeting of membrane fusion by SNARE mimicry: Convergent evolution ofLegionellaeffectors

Direct targeting of membrane fusion by SNARE mimicry: Convergent evolution ofLegionellaeffectors

PNA‐Hybridsequenzen als Erkennungseinheiten in SNARE‐Protein‐analogen Peptiden

Supramolecular Control over Split‐Luciferase Complementation

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