Abstract
Cryo-electron microscopy (cryo-EM) can solve structures of highly dynamic macromolecular complexes. To characterize less well defined regions in cryo-EM images, cross-linking coupled with mass spectrometry (CX-MS) provides valuable information on the arrangement of domains and amino acids. CX-MS involves covalent linkage of protein residues close to each other and identifying these connections by mass spectrometry. Here, we summarise the advances of CX-MS and its integration with cryo-EM for structural reconstruction.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Literatur
Cheng Y (2015) Single-particle cryo-EM at crystallographic resolution. Cell 161:450–457
Cheng Y, Grigorieff N, Penczek PA et al. (2015) A primer to single-particle cryo-electron microscopy. Cell 161:438–449
Nogales E, Scheres SH (2015) Cryo-EM: a unique tool for the visualization of macromolecular complexity. Mol Cell 58:677–689
Fernandez-Leiro R, Scheres SH (2016) Unravelling biological macromolecules with cryo-electron microscopy. Nature 537:339–346
Schmidt C, Urlaub H (2017) Combining cryo-electron microscopy (cryo-EM) and cross-linking mass spectrometry (CX-MS) for structural elucidation of large protein assemblies. Curr Opin Struct Biol 46:157–168
Sinz A (2003) Chemical cross-linking and mass spectrometry for mapping three-dimensional structures of proteins and protein complexes. J Mass Spectrom 38:1225–1237
Rappsilber J (2011) The beginning of a beautiful friendship: cross-linking/mass spectrometry and modelling of proteins and multi-protein complexes. J Struct Biol 173:530–540
Leitner A, Faini M, Stengel F et al. (2016) Crosslinking and mass spectrometry: an integrated technology to understand the structure and function of molecular machines. Trends Biochem Sci 41:20–32
Kao A, Chiu CL, Vellucci D et al. (2011) Development of a novel cross-linking strategy for fast and accurate identification of cross-linked peptides of protein complexes. Mol Cell Proteomics 10, doi: 10.1074/mcp.M110.002212
Muller MQ, Dreiocker F, Ihling CH et al. (2010) Cleavable cross-linker for protein structure analysis: reliable identification of cross-linking products by tandem MS. Anal Chem 82:6958–6968
Liu F, Rijkers DT, Post H et al. (2015) Proteome-wide profiling of protein assemblies by cross-linking mass spectrometry. Nat Methods 12:1179–1184
Agafonov DE, Kastner B, Dybkov O et al. (2016) Molecular architecture of the human U4/U6.U5 tri-snRNP. Science 351:1416–1420
Rauhut R, Fabrizio P, Dybkov O et al. (2016) Molecular architecture of the Saccharomyces cerevisiae activated spliceosome. Science 353:1399–1405
Bertram K, Agafonov DE, Dybkov O et al. (2017) Cryo-EM structure of a pre-catalytic human spliceosome primed for activation. Cell 170:701–713
Bertram K, Agafonov DE, Liu WT et al. (2017) Cryo-EM structure of a human spliceosome activated for step 2 of splicing. Nature 542:318–323
Haselbach D, Komarov I, Agafonov DE et al. (2018) Structure and conformational dynamics of the human spliceosomal Bact complex. Cell 172:454–464
Author information
Authors and Affiliations
Corresponding author
Additional information
Das Fachgebiet Bioverfahrenstechnik der Technischen Universität Berlin wird seit 2008 von Prof. Dr. Peter Neubauer geleitet. Neben den Forschungsschwerpunkten Scale-up, prozessanalytische Technologien, Biokatalyse und Bioelektronik wird am Fachgebiet im Bereich der automatisierten Prozessentwicklung und Modellierung geforscht. Unter Leitung von Dr. M. Nicolas Cruz-Bournazou und Dr. Florian Glauche arbeitet ein interdisziplinäres Team an der Kombination von Laborautomatisierung und mathematischer Modellierung für eine zeit- und kosteneffiziente Charakterisierung zellulärer Wachstums- und Produktionsprozesse.
Rights and permissions
Open Access:
This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits use, duplication, adaption, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
Open access funding provided by Max Planck Society.
About this article
Cite this article
Dybkov, O., Stützer, A., Bertram, K. et al. Protein-Cross-Linking zur Aufklärung von komplexen Strukturen. Biospektrum 24, 278–282 (2018). https://doi.org/10.1007/s12268-018-0909-6
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12268-018-0909-6