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Chair of Biochemistry II

Cryo Electron Microscopy

The facility for electron cryo microscopy provides facilitated access to a Titan-Krios G3 with an X-FEG source, 300 kV, Selectris Energy filter and a Falcon IVi camera with direct electron detection. The set-up is optimized for high-resolution, high-throughput image acquisition for structure determination by single particle image analysis. Image acquisition can be in linear mode or in counting mode. The acquisition is automated using EPU  and can run over several days without further intervention. Typical, data acquisitions take 1-3 days. Longer sessions can also be accommodated if needed.

History

The facility started in 2018. In Jan 2023 support for single particle image processing (3 workstations) was added, which provides a compute environment for small processing projects (max. 6 weeks),  training and processing on behalf of users. In Sept 2023 the camera system was upgraded to a Falcon IVi with Selectris Energy Filter for faster high resolution acquisition in counting mode.

How to acknowledge?

Include the following statement into the acknowlwdgements of your publication:

“Electron Cryo Microscopy was carried out in the cryo EM-facility of the Julius-Maximilians-Universität Würzburg funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) – 359471283, 456578072, 525040890.”

Access and contact:

Access is fee-based and regulated by access rules, which can be downloaded here.

If you are interested in access , need a quote or want to discuss your project: Please contact cryo-em@uni-wuerzburg.de  

Publications that made use of the facility

M. Gerovac, K. Chihara, L. Wicke, B. Bottcher, R. Lavigne, J. Vogel (2024) Phage proteins target and co-opt host ribosomes immediately upon infection. Nat Microbiol 9: 787-800

S. Endres, S. Ehrmanntraut, L. Endres, K. Can, C. Kraft, T. Rasmussen, R. Luxenhofer, B. Bottcher, B. Engels and A. C. Poppler (2023) Structural Investigation on How Guest Loading of Poly(2-oxazoline)-Based Micelles Affects the Interaction with Simulated Intestinal Fluids. 10.1021/acsbiomaterials.3c00645

S. Sekulovski, L. Susac, L. S. Stelzl, R. Tampe and S. Trowitzsch (2023) Structural basis of substrate recognition by human tRNA splicing endonuclease TSEN. 30. 834-840.

S. Endres, E. Karaev, S. Hanio, J. Schlauersbach, C. Kraft, T. Rasmussen, R. Luxenhofer, B. Bottcher, L. Meinel and A. C. Poppler (2022)  Concentration and composition dependent aggregation of Pluronic- and Poly-(2-oxazolin)-Efavirenz formulations in biorelevant media. J Colloid Interface Sci  606, 1179-1192.

L. T. Alexander, R. Lepore, A. Kryshtafovych, A. Adamopoulos, M. Alahuhta, A. M. Arvin, Y. J. Bomble, B. Bottcher, C. Breyton, V. Chiarini, N. B. Chinnam, W. Chiu, K. Fidelis, R. Grinter, G. D. Gupta, M. D. Hartmann, C. S. Hayes, T. Heidebrecht, A. Ilari, A. Joachimiak, Y. Kim, R. Linares, A. L. Lovering, V. V. Lunin, A. N. Lupas, C. Makbul, K. Michalska, J. Moult, P. K. Mukherjee, W. S. Nutt, S. L. Oliver, A. Perrakis, L. Stols, J. A. Tainer, M. Topf, S. E. Tsutakawa, M. Valdivia-Delgado and T. Schwede (2021)  Target highlights in CASP14: Analysis of models by structure providers. Proteins  

V. J. Flegler, A. Rasmussen, K. Borbil, L. Boten, H. A. Chen, H. Deinlein, J. Halang, K. Hellmanzik, J. Loffler, V. Schmidt, C. Makbul, C. Kraft, R. Hedrich, T. Rasmussen and B. Bottcher (2021)  Mechanosensitive channel gating by delipidation. Proc Natl Acad Sci U S A  118, e2107095118.

A. Grauel, J. Kagi, T. Rasmussen, I. Makarchuk, S. Oppermann, A. F. A. Moumbock, D. Wohlwend, R. Muller, F. Melin, S. Gunther, P. Hellwig, B. Bottcher and T. Friedrich (2021)  Structure of Escherichia coli cytochrome bd-II type oxidase with bound aurachin D. Nat Commun  12, 6498.

C. Grimm, J. Bartuli, B. Boettcher, A. A. Szalay and U. Fischer (2021)  Structural basis of the complete poxvirus transcription initiation process. Nat Struct Mol Biol  28, 779-788.

F. B. Heiss, J. L. Daiss, P. Becker and C. Engel (2021)  Conserved strategies of RNA polymerase I hibernation and activation. Nat Commun  12, 758.

C. Makbul, V. Khayenko, H. M. Maric and B. Bottcher (2021)  Conformational Plasticity of Hepatitis B Core Protein Spikes Promotes Peptide Binding Independent of the Secretion Phenotype. Microorganisms  9, 956.

C. Makbul, C. Kraft, M. Grießmann, T. Rasmussen, K. Katzenberger, M. Lappe, P. Pfarr, C. Stoffer, M. Stöhr, A.-M. Wandinger and B. Böttcher (2021)  Binding of a Pocket Factor to Hepatitis B Virus Capsids Changes the Rotamer Conformation of Phenylalanine 97. Viruses  13, 2115.

V. J. Flegler, A. Rasmussen, S. Rao, N. Wu, R. Zenobi, M. S. P. Sansom, R. Hedrich, T. Rasmussen and B. Bottcher (2020)  The MscS-like channel YnaI has a gating mechanism based on flexible pore helices. Proc Natl Acad Sci U S A  117, 28754-28762.

C. Makbul, M. Nassal and B. Bottcher (2020)  Slowly folding surface extension in the prototypic avian hepatitis B virus capsid governs stability. Elife  9, e57277.

K. Stokes, A. Winczura, B. Song, G. Piccoli and D. B. Grabarczyk (2020)  Ctf18-RFC and DNA Pol form a stable leading strand polymerase/clamp loader complex required for normal and perturbed DNA replication. Nucleic Acids Res  48, 8128-8145.

J. Wiest, J. Kehrein, M. Saedtler, K. Schilling, E. Cataldi, C. A. Sotriffer, U. Holzgrabe, T. Rasmussen, B. Bottcher, M. Cronin-Golomb, M. Lehmann, N. Jung, M. Windbergs and L. Meinel (2020)  Controlling Supramolecular Structures of Drugs by Light. Mol Pharm  17, 4704-4708.

N. Famelis, A. Rivera-Calzada, G. Degliesposti, M. Wingender, N. Mietrach, J. M. Skehel, R. Fernandez-Leiro, B. Böttcher, A. Schlosser, O. Llorca and S. Geibel (2019)  Architecture of the mycobacterial type VII secretion system. Nature  576, 321-325.

C. Grimm, H. S. Hillen, K. Bedenk, J. Bartuli, S. Neyer, Q. Zhang, A. Huttenhofer, M. Erlacher, C. Dienemann, A. Schlosser, H. Urlaub, B. Böttcher, A. A. Szalay, P. Cramer and U. Fischer (2019)  Structural Basis of Poxvirus Transcription: Vaccinia RNA Polymerase Complexes. Cell  179, 1537-1550 e19.

T. Rasmussen, V. J. Flegler, A. Rasmussen and B. Böttcher (2019)  Structure of the Mechanosensitive Channel MscS Embedded in the Membrane Bilayer. J Mol Biol  431, 3081-3090.

B. Song, J. Lenhart, V. J. Flegler, C. Makbul, T. Rasmussen and B. Böttcher (2019)  Capabilities of the Falcon III detector for single-particle structure determination. Ultramicroscopy  203, 145-154.

A. Thesseling, T. Rasmussen, S. Burschel, D. Wohlwend, J. Kagi, R. Muller, B. Böttcher and T. Friedrich (2019)  Homologous bd oxidases share the same architecture but differ in mechanism. Nat Commun  10, 5138.