Jahn, Reinhard

[["dc.bibliographiccitation.firstpage","2858"],["dc.bibliographiccitation.issue","9"],["dc.bibliographiccitation.journal","Proceedings of the National Academy of Sciences"],["dc.bibliographiccitation.lastpage","2863"],["dc.bibliographiccitation.volume","101"],["dc.contributor.author","Schuette, C. G."],["dc.contributor.author","Hatsuzawa, K."],["dc.contributor.author","Margittai, M."],["dc.contributor.author","Stein, A."],["dc.contributor.author","Riedel, D."],["dc.contributor.author","Kuster, P."],["dc.contributor.author","Konig, M."],["dc.contributor.author","Seidel, C."],["dc.contributor.author","Jahn, R."],["dc.date.accessioned","2021-06-01T10:51:03Z"],["dc.date.available","2021-06-01T10:51:03Z"],["dc.date.issued","2004"],["dc.identifier.doi","10.1073/pnas.0400044101"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/86875"],["dc.language.iso","en"],["dc.notes.intern","DOI-Import GROB-425"],["dc.relation.eissn","1091-6490"],["dc.relation.issn","0027-8424"],["dc.title","Determinants of liposome fusion mediated by synaptic SNARE proteins"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","606"],["dc.bibliographiccitation.issue","6"],["dc.bibliographiccitation.journal","Microscopy research and technique"],["dc.bibliographiccitation.lastpage","617"],["dc.bibliographiccitation.volume","73"],["dc.contributor.author","Geumann, Ulf"],["dc.contributor.author","Schaefer, Christina"],["dc.contributor.author","Riedel, Dietmar"],["dc.contributor.author","Jahn, Reinhard"],["dc.contributor.author","Rizzoli, Silvio"],["dc.date.accessioned","2017-09-07T11:46:02Z"],["dc.date.available","2017-09-07T11:46:02Z"],["dc.date.issued","2010"],["dc.description.abstract","In the plasma membrane, membrane proteins are frequently organized in microdomains that are stabilized both by protein-protein and protein-lipid interactions, with the membrane lipid cholesterol being instrumental for microdomain stability. However, it is unclear whether such microdomains persist during endocytotic membrane trafficking. We used stimulated emission-depletion microscopy to investigate the domain structure of the endosomes. We developed a semiautomatic method for counting the individual domains, an approach that we have validated by immunoelectron microscopy. We found that in endosomes derived from neuroendocrine PC12 cells synaptophysin and several SNARE proteins are organized in microdomains. Cholesterol depletion by methyl-beta-cyclodextrin disintegrates most of the domains. Interestingly, no change in the frequency of microdomains was observed when endosomes were fused with protein-free liposomes of similar size (in what constitutes a novel approach in modifying acutely the lipid composition of organelles), regardless of whether the membrane lipid composition of the liposomes was similar or very different from that of the endosomes. Similarly, Rab depletion from the endosome membranes left the domain structure unaffected. Furthermore, labeled exogenous protein, introduced into endosomes by liposome fusion, equilibrated with the corresponding microdomains. We conclude that synaptic membrane proteins are organized in stable but dynamic clusters within endosomes, which are likely to persist during membrane recycling. Microsc. Res. Tech. 73:606-617, 2010. (C) 2009 Wiley-Liss, Inc."],["dc.identifier.doi","10.1002/jemt.20800"],["dc.identifier.gro","3142915"],["dc.identifier.isi","000278641200004"],["dc.identifier.pmid","19937745"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/372"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.publisher","Wiley-liss"],["dc.relation.issn","1059-910X"],["dc.title","Synaptic Membrane Proteins Form Stable Microdomains in Early Endosomes"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","805"],["dc.bibliographiccitation.issue","7"],["dc.bibliographiccitation.journal","Nature Structural & Molecular Biology"],["dc.bibliographiccitation.lastpage","U82"],["dc.bibliographiccitation.volume","18"],["dc.contributor.author","van den Bogaart, Geert"],["dc.contributor.author","Thutupalli, Shashi"],["dc.contributor.author","Risselada, J. H."],["dc.contributor.author","Meyenberg, Karsten"],["dc.contributor.author","Holt, Matthew"],["dc.contributor.author","Riedel, Dietmar"],["dc.contributor.author","Diederichsen, Ulf"],["dc.contributor.author","Herminghaus, Stephan"],["dc.contributor.author","Grubmüller, Helmut"],["dc.contributor.author","Jahn, Reinhard"],["dc.date.accessioned","2017-09-07T11:44:10Z"],["dc.date.available","2017-09-07T11:44:10Z"],["dc.date.issued","2011"],["dc.description.abstract","Synaptotagmin-1 triggers Ca2+-sensitive, rapid neurotransmitter release by promoting interactions between SNARE proteins on synaptic vesicles and the plasma membrane. How synaptotagmin-1 promotes this interaction is unclear, and the massive increase in membrane fusion efficiency of Ca2+-bound synaptotagmin-1 has not been reproduced in vitro. However, previous experiments have been performed at relatively high salt concentrations, screening potentially important electrostatic interactions. Using functional reconstitution in liposomes, we show here that at low ionic strength SNARE-mediated membrane fusion becomes strictly dependent on both Ca2+ and synaptotagmin-1. Under these conditions, synaptotagmin-1 functions as a distance regulator that tethers the liposomes too far from the plasma membrane for SNARE nucleation in the absence of Ca2+, but while bringing the liposomes close enough for membrane fusion in the presence of Ca2+. These results may explain how the relatively weak electrostatic interactions between synaptotagmin-1 and membranes substantially accelerate fusion."],["dc.identifier.doi","10.1038/nsmb.2061"],["dc.identifier.gro","3142704"],["dc.identifier.isi","000292507500009"],["dc.identifier.pmid","21642968"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/138"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","1545-9993"],["dc.title","Synaptotagmin-1 may be a distance regulator acting upstream of SNARE nucleation"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","13441"],["dc.bibliographiccitation.issue","40"],["dc.bibliographiccitation.journal","The Journal of neuroscience"],["dc.bibliographiccitation.lastpage","13453"],["dc.bibliographiccitation.volume","30"],["dc.contributor.author","Pavlos, Nathan J."],["dc.contributor.author","Gronborg, Mads"],["dc.contributor.author","Riedel, Dietmar"],["dc.contributor.author","Chua, John Jia En"],["dc.contributor.author","Boyken, Janina"],["dc.contributor.author","Kloepper, Tobias H."],["dc.contributor.author","Urlaub, Henning"],["dc.contributor.author","Rizzoli, Silvio"],["dc.contributor.author","Jahn, Reinhard"],["dc.date.accessioned","2017-09-07T11:45:15Z"],["dc.date.available","2017-09-07T11:45:15Z"],["dc.date.issued","2010"],["dc.description.abstract","Rab GTPases are molecular switches that orchestrate protein complexes before membrane fusion reactions. In synapses, Rab3 and Rab5 proteins have been implicated in the exo-endocytic cycling of synaptic vesicles (SVs), but an involvement of additional Rabs cannot be excluded. Here, combining high-resolution mass spectrometry and chemical labeling (iTRAQ) together with quantitative immunoblotting and fluorescence microscopy, we have determined the exocytotic (Rab3a, Rab3b, Rab3c, and Rab27b) and endocytic (Rab4b, Rab5a/b, Rab10, Rab11b, and Rab14) Rab machinery of SVs. Analysis of two closely related proteins, Rab3a and Rab27b, revealed colocalization in synaptic nerve terminals, where they reside on distinct but overlapping SV pools. Moreover, whereas Rab3a readily dissociates from SVs during Ca2+-triggered exocytosis, and is susceptible to membrane extraction by Rab-GDI, Rab27b persists on SV membranes upon stimulation and is resistant to GDI-coupled Rab retrieval. Finally, we demonstrate that selective modulation of the GTP/GDP switch mechanism of Rab27b impairs SV recycling, suggesting that Rab27b, probably in concert with Rab3s, is involved in SV exocytosis."],["dc.identifier.doi","10.1523/JNEUROSCI.0907-10.2010"],["dc.identifier.gro","3142843"],["dc.identifier.isi","000282571000024"],["dc.identifier.pmid","20926670"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/292"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.publisher","Soc Neuroscience"],["dc.relation.issn","0270-6474"],["dc.title","Quantitative Analysis of Synaptic Vesicle Rabs Uncovers Distinct Yet Overlapping Roles for Rab3a and Rab27b in Ca2+-Triggered Exocytosis"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","831"],["dc.bibliographiccitation.issue","4"],["dc.bibliographiccitation.journal","Cell"],["dc.bibliographiccitation.lastpage","846"],["dc.bibliographiccitation.volume","127"],["dc.contributor.author","Takamori, Shigeo"],["dc.contributor.author","Holt, Matthew"],["dc.contributor.author","Stenius, Katinka"],["dc.contributor.author","Lemke, Edward A."],["dc.contributor.author","Gronborg, Mads"],["dc.contributor.author","Riedel, Dietmar"],["dc.contributor.author","Urlaub, Henning"],["dc.contributor.author","Schenck, Stephan"],["dc.contributor.author","Brügger, Britta"],["dc.contributor.author","Ringler, Philippe"],["dc.contributor.author","Müller, Shirley A."],["dc.contributor.author","Rammner, Burkhard"],["dc.contributor.author","Graeter, Frauke"],["dc.contributor.author","Hub, Jochen S."],["dc.contributor.author","Groot, Bert L. de"],["dc.contributor.author","Mieskes, Gottfried"],["dc.contributor.author","Moriyama, Yoshinori"],["dc.contributor.author","Klingauf, Juergen"],["dc.contributor.author","Grubmüller, Helmut"],["dc.contributor.author","Heuser, John"],["dc.contributor.author","Wieland, Felix"],["dc.contributor.author","Jahn, Reinhard"],["dc.date.accessioned","2017-09-07T11:49:54Z"],["dc.date.available","2017-09-07T11:49:54Z"],["dc.date.issued","2006"],["dc.description.abstract","Membrane traffic in eukaryotic cells involves transport of vesicles that bud from a donor compartment and fuse with an acceptor compartment. Common principles of budding and fusion have emerged, and many of the proteins involved in these events are now known. However, a detailed picture of an entire trafficking organelle is not yet available. Using synaptic vesicles as a model, we have now determined the protein and lipid composition; measured vesicle size, density, and mass; calculated the average protein and lipid mass per vesicle; and determined the copy number of more than a dozen major constituents. A model has been constructed that integrates all quantitative data and includes structural models of abundant proteins. Synaptic vesicles are dominated by proteins, possess a surprising diversity of trafficking proteins, and, with the exception of the V-ATPase that is present in only one to two copies, contain numerous copies of proteins essential for membrane traffic and neurotransmitter uptake."],["dc.identifier.doi","10.1016/j.cell.2006.10.030"],["dc.identifier.gro","3143587"],["dc.identifier.isi","000242330600027"],["dc.identifier.pmid","17110340"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/1117"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","0092-8674"],["dc.title","Molecular anatomy of a trafficking organelle"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","1200"],["dc.bibliographiccitation.issue","7"],["dc.bibliographiccitation.journal","Biophysical Journal"],["dc.bibliographiccitation.lastpage","1208"],["dc.bibliographiccitation.volume","98"],["dc.contributor.author","Castorph, Simon"],["dc.contributor.author","Riedel, Dietmar"],["dc.contributor.author","Arleth, Lise"],["dc.contributor.author","Sztucki, Michael"],["dc.contributor.author","Jahn, Reinhard"],["dc.contributor.author","Holt, Matthew"],["dc.contributor.author","Salditt, Tim"],["dc.date.accessioned","2017-09-07T11:46:06Z"],["dc.date.available","2017-09-07T11:46:06Z"],["dc.date.issued","2010"],["dc.description.abstract","Synaptic vesicles (SVs) are small, membrane-bound organelles that are found in the synaptic terminal of neurons, and which are crucial in neurotransmission. After a rise in internal [Ca2+] during neuronal stimulation, SVs fuse with the plasma membrane releasing their neurotransmitter content, which then signals neighboring neurons. SVs are subsequently recycled and refilled with neurotransmitter for further rounds of release. Recently, tremendous progress has been made in elucidating the molecular composition of SVs, as well as putative protein-protein interactions. However, what is lacking is an empirical description of SV structure at the supramolecular level which is necessary to enable us to fully understand the processes of membrane fusion, retrieval, and recycling. Using small-angle x-ray scattering, we have directly investigated the size and structure of purified SVs. From this information, we deduced detailed size and density parameters for the protein layers responsible for SV function, as well as information about the lipid bilayer. To achieve a convincing model fit, a laterally anisotropic structure for the protein shell is needed, as a rotationally symmetric density profile does not explain the data. Not only does our model confirm many of the preexisting ideas concerning SV structure, but also for the first time, to our knowledge, it indicates structural refinements, such as the presence of protein microdomains."],["dc.identifier.doi","10.1016/j.bpj.2009.12.4278"],["dc.identifier.gro","3142938"],["dc.identifier.isi","000276582700012"],["dc.identifier.pmid","20371319"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/397"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","0006-3495"],["dc.relation.orgunit","Institut für Röntgenphysik"],["dc.relation.workinggroup","RG Salditt (Structure of Biomolecular Assemblies and X-Ray Physics)"],["dc.subject.gro","x-ray scattering"],["dc.subject.gro","membrane biophysics"],["dc.title","Structure Parameters of Synaptic Vesicles Quantified by Small-Angle X-Ray Scattering"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original_ja"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.artnumber","e05597"],["dc.bibliographiccitation.journal","eLife"],["dc.bibliographiccitation.volume","4"],["dc.contributor.author","Binotti, Beyenech"],["dc.contributor.author","Pavlos, Nathan J."],["dc.contributor.author","Riedel, Dietmar"],["dc.contributor.author","Wenzel, Dirk"],["dc.contributor.author","Vorbrueggen, Gerd"],["dc.contributor.author","Schalk, Amanda M."],["dc.contributor.author","Kuehnel, Karin"],["dc.contributor.author","Boyken, Janina"],["dc.contributor.author","Erck, Christian"],["dc.contributor.author","Martens, Henrik"],["dc.contributor.author","Chua, John J. E."],["dc.contributor.author","Jahn, Reinhard"],["dc.date.accessioned","2018-11-07T10:01:07Z"],["dc.date.available","2018-11-07T10:01:07Z"],["dc.date.issued","2015"],["dc.description.abstract","Small GTPases of the Rab family not only regulate target recognition in membrane traffic but also control other cellular functions such as cytoskeletal transport and autophagy. Here we show that Rab26 is specifically associated with clusters of synaptic vesicles in neurites. Overexpression of active but not of GDP-preferring Rab26 enhances vesicle clustering, which is particularly conspicuous for the EGFP-tagged variant, resulting in a massive accumulation of synaptic vesicles in neuronal somata without altering the distribution of other organelles. Both endogenous and induced clusters co-localize with autophagy-related proteins such as Atg16L1, LC3B and Rab33B but not with other organelles. Furthermore, Atg16L1 appears to be a direct effector of Rab26 and binds Rab26 in its GTP-bound form, albeit only with low affinity. We propose that Rab26 selectively directs synaptic and secretory vesicles into preautophagosomal structures, suggesting the presence of a novel pathway for degradation of synaptic vesicles."],["dc.identifier.doi","10.7554/eLife.05597"],["dc.identifier.isi","000348683800001"],["dc.identifier.pmid","25643395"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/37947"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Elife Sciences Publications Ltd"],["dc.relation.issn","2050-084X"],["dc.title","The GTPase Rab26 links synaptic vesicles to the autophagy pathway"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","981"],["dc.bibliographiccitation.issue","6276"],["dc.bibliographiccitation.journal","Science"],["dc.bibliographiccitation.lastpage","984"],["dc.bibliographiccitation.volume","351"],["dc.contributor.author","Farsi, Zohreh"],["dc.contributor.author","Preobraschenski, Julia"],["dc.contributor.author","van den Bogaart, Geert"],["dc.contributor.author","Riedel, Dietmar"],["dc.contributor.author","Jahn, Reinhard"],["dc.contributor.author","Woehler, Andrew"],["dc.date.accessioned","2019-12-03T15:05:23Z"],["dc.date.available","2019-12-03T15:05:23Z"],["dc.date.issued","2016"],["dc.description.abstract","Synaptic transmission is mediated by the release of neurotransmitters, which involves exo-endocytotic cycling of synaptic vesicles. To maintain synaptic function, synaptic vesicles are refilled with thousands of neurotransmitter molecules within seconds after endocytosis, using the energy provided by an electrochemical proton gradient. However, it is unclear how transmitter molecules carrying different net charges can be efficiently sequestered while maintaining charge neutrality and osmotic balance. We used single-vesicle imaging to monitor pH and electrical gradients and directly showed different uptake mechanisms for glutamate and γ-aminobutyric acid (GABA) operating in parallel. In contrast to glutamate, GABA was exchanged for protons, with no other ions participating in the transport cycle. Thus, only a few components are needed to guarantee reliable vesicle filling with different neurotransmitters."],["dc.identifier.doi","10.1126/science.aad8142"],["dc.identifier.pmid","26912364"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/62729"],["dc.language.iso","en"],["dc.relation.eissn","1095-9203"],["dc.relation.issn","0036-8075"],["dc.relation.issn","1095-9203"],["dc.title","Single-vesicle imaging reveals different transport mechanisms between glutamatergic and GABAergic vesicles"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","358"],["dc.bibliographiccitation.issue","3"],["dc.bibliographiccitation.journal","Nature Structural & Molecular Biology"],["dc.bibliographiccitation.lastpage","U129"],["dc.bibliographiccitation.volume","17"],["dc.contributor.author","van den Bogaart, Geert"],["dc.contributor.author","Holt, Matthew G."],["dc.contributor.author","Bunt, Gertrude"],["dc.contributor.author","Riedel, Dietmar"],["dc.contributor.author","Wouters, Fred S."],["dc.contributor.author","Jahn, Reinhard"],["dc.date.accessioned","2018-11-07T08:45:38Z"],["dc.date.available","2018-11-07T08:45:38Z"],["dc.date.issued","2010"],["dc.description.abstract","In eukaryotes, most intracellular membrane fusion reactions are mediated by the interaction of SNARE proteins that are present in both fusing membranes. However, the minimal number of SNARE complexes needed for membrane fusion is not known. Here we show unambiguously that one SNARE complex is sufficient for membrane fusion. We performed controlled in vitro Forster resonance energy transfer (FRET) experiments and found that liposomes bearing only a single SNARE molecule are still capable of fusion with other liposomes or with purified synaptic vesicles. Furthermore, we demonstrated that multiple SNARE complexes do not act cooperatively, showing that synergy between several SNARE complexes is not needed for membrane fusion. Our findings shed new light on the mechanism of SNARE-mediated membrane fusion and call for a revision of current views of fusion events such as the fast release of neurotransmitters."],["dc.identifier.doi","10.1038/nsmb.1748"],["dc.identifier.isi","000275182700017"],["dc.identifier.pmid","20139985"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/20493"],["dc.notes.status","zu prüfen"],["dc.notes.submitter","Najko"],["dc.publisher","Nature Publishing Group"],["dc.relation.issn","1545-9993"],["dc.title","One SNARE complex is sufficient for membrane fusion"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.status","published"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","6000"],["dc.bibliographiccitation.issue","22"],["dc.bibliographiccitation.journal","EMBO Journal"],["dc.bibliographiccitation.lastpage","6010"],["dc.bibliographiccitation.volume","19"],["dc.contributor.author","Ossig, R."],["dc.contributor.author","Schmitt, H. D."],["dc.contributor.author","de Groot, Bert L."],["dc.contributor.author","Riedel, Dietmar"],["dc.contributor.author","Keränen, S."],["dc.contributor.author","Ronne, H."],["dc.contributor.author","Grubmüller, Helmut"],["dc.contributor.author","Jahn, Reinhard"],["dc.date.accessioned","2017-09-07T11:46:44Z"],["dc.date.available","2017-09-07T11:46:44Z"],["dc.date.issued","2000"],["dc.description.abstract","Assembly of SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors) mediates membrane fusions in all eukaryotic cells. The synaptic SNARE complex is represented by a twisted bundle of four a-helices, Leucine zipper-like layers extend through the length of the complex except for an asymmetric and ionic middle layer formed by three glutamines (Q) and one arginine (R), We have examined the functional consequences of Q-R exchanges in the conserved middle layer using the exocytotic SNAREs of yeast as a model. Exchanging Q for R in Sso2p drastically reduces cell growth and protein secretion. When a 3Q/1R ratio is restored by a mirror R-->Q substitution in the R-SNARE Snc2p, wild-type functionality is observed. Secretion is near normal when all four helices contain Q, but defects become apparent when additional mutations are present in other layers. Using molecular dynamics free energy perturbation simulations, these findings are rationalized in structural and energetic terms. We conclude that the asymmetric arrangement of the polar amino acids in the central layer is essential for normal function of SNAREs in membrane fusion."],["dc.description.abstract","Assembly of SNAREs (soluble N‐ethylmaleimide‐ sensitive factor attachment protein receptors) mediates membrane fusions in all eukaryotic cells. The synaptic SNARE complex is represented by a twisted bundle of four α‐helices. Leucine zipper‐like layers extend through the length of the complex except for an asymmetric and ionic middle layer formed by three glutamines (Q) and one arginine (R). We have examined the functional consequences of Q–R exchanges in the conserved middle layer using the exocytotic SNAREs of yeast as a model. Exchanging Q for R in Sso2p drastically reduces cell growth and protein secretion. When a 3Q/1R ratio is restored by a mirror R→Q substitution in the R‐SNARE Snc2p, wild‐type functionality is observed. Secretion is near normal when all four helices contain Q, but defects become apparent when additional mutations are present in other layers. Using molecular dynamics free energy perturbation simulations, these findings are rationalized in structural and energetic terms. We conclude that the asymmetric arrangement of the polar amino acids in the central layer is essential for normal function of SNAREs in membrane fusion."],["dc.identifier.doi","10.1093/emboj/19.22.6000"],["dc.identifier.gro","3144341"],["dc.identifier.isi","000165558900007"],["dc.identifier.pmid","11080147"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/1954"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","0261-4189"],["dc.title","Exocytosis requires asymmetry in the central layer of the SNARE complex"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]