potassium channel; 2D crystals; membrane protein structure; electron crystallography; electron tomography
Abeyrathne Priyanka D, Chami Mohamed, Stahlberg Henning (2016), Biochemical and biophysical approaches to study the structure and function of the chloride channel (ClC) family of proteins., in Biochimie
, 128-129, 154-62.
Lauer Matthias E, Graff-Meyer Alexandra, Rufer Arne C, Maugeais Cyrille, von der Mark Elisabeth, Matile Hugues, D'Arcy Brigitte, Magg Christine, Ringler Philippe, Müller Shirley A, Scherer Sebastian, Dernick Gregor, Thoma Ralf, Hennig Michael, Niesor Eric J, Stahlberg Henning (2016), Cholesteryl ester transfer between lipoproteins does not require a ternary tunnel complex with CETP., in Journal of structural biology
, 194(2), 191-8.
Castaño-Díez Daniel, Kudryashev Mikhail, Stahlberg Henning (2016), Dynamo Catalogue: Geometrical tools and data management for particle picking in subtomogram averaging of cryo-electron tomograms., in Journal of structural biology
Sborgi Lorenzo, Rühl Sebastian, Mulvihill Estefania, Pipercevic Joka, Heilig Rosalie, Stahlberg Henning, Farady Christopher J, Müller Daniel J, Broz Petr, Hiller Sebastian (2016), GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death., in The EMBO journal
, 35(16), 1766-78.
Taylor Nicholas M I, Prokhorov Nikolai S, Guerrero-Ferreira Ricardo C, Shneider Mikhail M, Browning Christopher, Goldie Kenneth N, Stahlberg Henning, Leiman Petr G (2016), Structure of the T4 baseplate and its function in triggering sheath contraction., in Nature
, 533(7603), 346-52.
Ayciriex Sophie, Gerber Hermeto, Osuna Guillermo M Garcia, Chami Mohamed, Stahlberg Henning, Shevchenko Andrej, Fraering Patrick C (2016), The lipidome associated with the γ-secretase complex is required for its integrity and activity., in The Biochemical journal
, 473(3), 321-34.
Kudryashev Mikhail, Castaño-Díez Daniel, Deluz Cédric, Hassaine Gherici, Grasso Luigino, Graf-Meyer Alexandra, Vogel Horst, Stahlberg Henning (2016), The Structure of the Mouse Serotonin 5-HT3 Receptor in Lipid Vesicles., in Structure (London, England : 1993)
, 24(1), 165-70.
Marabini Roberto, Carragher Bridget, Chen Shaoxia, Chen James, Cheng Anchi, Downing Kenneth H, Frank Joachim, Grassucci Robert A, Bernard Heymann J, Jiang Wen, Jonic Slavica, Liao Hstau Y, Ludtke Steven J, Patwari Shail, Piotrowski Angela L, Quintana Adrian, Sorzano Carlos O S, Stahlberg Henning, Vargas Javier, Voss Neil R, Chiu Wah, Carazo Jose M (2015), CTF Challenge: Result summary., in Journal of structural biology
, 190(3), 348-59.
Kudryashev Mikhail, Wang Ray Yu-Ruei, Brackmann Maximilian, Scherer Sebastian, Maier Timm, Baker David, DiMaio Frank, Stahlberg Henning, Egelman Edward H, Basler Marek (2015), Structure of the type VI secretion system contractile sheath., in Cell
, 160(5), 952-62.
Kudryashev Mikhail, Diepold Andreas, Amstutz Marlise, Armitage Judith P, Stahlberg Henning, Cornelis Guy R (2015), Yersinia enterocolitica type III secretion injectisomes form regularly spaced clusters, which incorporate new machines upon activation., in Molecular microbiology
, 95(5), 875-84.
McCoy Jason G, Rusinova Radda, Kim Dorothy M, Kowal Julia, Banerjee Sourabh, Jaramillo Cartagena Alexis, Thompson Ameer N, Kolmakova-Partensky Ludmila, Stahlberg Henning, Andersen Olaf S, Nimigean Crina M (2014), A KcsA/MloK1 chimeric ion channel has lipid-dependent ligand-binding energetics., in The Journal of biological chemistry
, 289(14), 9535-46.
Goldie Kenneth N, Abeyrathne Priyanka, Kebbel Fabian, Chami Mohamed, Ringler Philippe, Stahlberg Henning (2014), Cryo-electron microscopy of membrane proteins., in Methods in molecular biology (Clifton, N.J.)
, 1117, 325-41.
Kowal Justyna Ł, Kowal Julia K, Wu Dalin, Stahlberg Henning, Palivan Cornelia G, Meier Wolfgang P (2014), Functional surface engineering by nucleotide-modulated potassium channel insertion into polymer membranes attached to solid supports., in Biomaterials
, 35(26), 7286-94.
Kowal Julia, Chami Mohamed, Baumgartner Paul, Arheit Marcel, Chiu Po-Lin, Rangl Martina, Scheuring Simon, Schröder Gunnar F, Nimigean Crina M, Stahlberg Henning (2014), Ligand-induced structural changes in the cyclic nucleotide-modulated potassium channel MloK1., in Nature communications
, 5, 3106-3106.
Kudryashev Misha, Aktoudianaki Aikaterini, Dedoglou Dimitrios, Stahlberg Henning, Tsiotis Georgios (2014), The ultrastructure of Chlorobaculum tepidum revealed by cryo-electron tomography., in Biochimica et biophysica acta
, 1837(10), 1635-42.
Hassaine Ghérici, Deluz Cédric, Grasso Luigino, Wyss Romain, Tol Menno B, Hovius Ruud, Graff Alexandra, Stahlberg Henning, Tomizaki Takashi, Desmyter Aline, Moreau Christophe, Li Xiao-Dan, Poitevin Frédéric, Vogel Horst, Nury Hugues (2014), X-ray structure of the mouse serotonin 5-HT3 receptor., in Nature
, 512(7514), 276-81.
Kudryashev Mikhail, Stenta Marco, Schmelz Stefan, Amstutz Marlise, Wiesand Ulrich, Castaño-Díez Daniel, Degiacomi Matteo T, Münnich Stefan, Bleck Christopher Ke, Kowal Julia, Diepold Andreas, Heinz Dirk W, Dal Peraro Matteo, Cornelis Guy R, Stahlberg Henning (2013), In situ structural analysis of the Yersinia enterocolitica injectisome., in eLife
, 2, 00792-00792.
Kebbel Fabian, Kurz Mareike, Arheit Marcel, Grütter Markus G, Stahlberg Henning (2013), Structure and substrate-induced conformational changes of the secondary citrate/sodium symporter CitS revealed by electron crystallography., in Structure (London, England : 1993)
, 21(7), 1243-50.
Kowal Julia, Chami Mohamed, Ringler Philippe, Müller Shirley A, Kudryashev Mikhail, Castaño-Díez Daniel, Amstutz Marlise, Cornelis Guy R, Stahlberg Henning, Engel Andreas (2013), Structure of the dodecameric Yersinia enterocolitica secretin YscC and its trypsin-resistant core., in Structure (London, England : 1993)
, 21(12), 2152-61.
Tol Menno B, Deluz Cédric, Hassaine Gherici, Graff Alexandra, Stahlberg Henning, Vogel Horst (2013), Thermal unfolding of a mammalian pentameric ligand-gated ion channel proceeds at consecutive, distinct steps., in The Journal of biological chemistry
, 288(8), 5756-69.
Membrane proteins are central to health and disease. A detailed understanding of the function of membrane proteins requires access to high-resolution structural data and mechanistic insight into conformational changes occurring when the proteins are active. We here propose to employ the recently established cryo-electron microscopy imaging capabilities in C-CINA, the Center for Cellular Imaging and NanoAnalytics, and our newly developed image processing software, to study the dynamic structure of selected membrane protein systems in liposomes, a close-to-native environment: Vesicles containing excellently ordered two-dimensional (2D) crystalline membrane protein arrays will be vitrified in aqueous buffer solutions, whereby different buffer solutions will be established on the inside and outside of the liposomes. The resulting buffer gradient will impose an electric potential, pH and/or ligand concentration gradient across the membranes, which will be experienced by the reconstituted proteins. Further, 2D crystals grown with a 180-degree screw-axis symmetry will place adjacent membrane proteins in alternating orientations in the membranes, allowing us to determine the 3D structures of oppositely regulated states in one experiment.We will employ a combination of electron crystallography (using our 2dx software), electron tomography (using our Dynamo software), and single particle processing of 2D crystals (using our new module in the 2dx software), to determine the 3D structure at 7Å resolution or better of selected systems. We already have excellently ordered 2D crystals of the bacterial iron-ferrichrome transporter FhuA decorated with the bacteriophage T5 tip protein pb5, of the cAMP-modulated potassium channel MloK1 from M. loti, and of the sodium/citrate symporter CitS from K. pneumonia, and we will grow 2D crystals of the voltage gated potassium channel KvAP from A. pernix. The exceptional quality and contrast of the FhuA-pb5 crystals will yield a high-resolution structure and allow our new methodology to be finely tuned. Analysis of the CitS crystals will reveal the active site and document a new, unknown fold of an important class of membrane proteins. We will expose MloK1 crystals to varying cAMP concentrations on both sides of the lipid membrane, and KvAP crystals to a Nernst electrical potential difference to trigger voltage gating, and document the conformational changes. The produced data will complement existing high-resolution X-ray structural studies with information about the dynamic functioning of these classes of membrane proteins. Importantly, this work will, for the first time, allow the direct structural observation of the voltage gating mechanism of a voltage-gated potassium channel.