title: Blind testing of cross-linking/mass spectrometry hybrid methods in CASP11.
abstract: Hybrid approaches combine computational methods with experimental data. The information contained in the experimental data can be leveraged to probe the structure of proteins otherwise elusive to computational methods. Compared with computational methods, the structures produced by hybrid methods exhibit some degree of experimental validation. In spite of these advantages, most hybrid methods have not yet been validated in blind tests, hampering their development. Here, we describe the first blind test of a specific cross-link based hybrid method in CASP. This blind test was coordinated by the CASP organizers and utilized a novel, high-density cross-linking/mass-spectrometry (CLMS) approach that is able to collect high-density CLMS data in a matter of days. This experimental protocol was developed in the Rappsilber laboratory. This approach exploits the chemistry of a highly reactive, photoactivatable cross-linker to produce an order of magnitude more cross-links than homobifunctional cross-linkers. The Rappsilber laboratory generated experimental CLMS data based on this protocol, submitted the data to the CASP organizers which then released this data to the CASP11 prediction groups in a separate, CLMS assisted modeling experiment. We did not observe a clear improvement of assisted models, presumably because the properties of the CLMS data-uncertainty in cross-link identification and residue-residue assignment, and uneven distribution over the protein-were largely unknown to the prediction groups and their approaches were not yet tailored to this kind of data. We also suggest modifications to the CLMS-CASP experiment and discuss the importance of rigorous blind testing in the development of hybrid methods. Proteins 2016; 84(Suppl 1):152-163. © 2016 The Authors Proteins: Structure, Function, and Bioinformatics Published by Wiley Periodicals, Inc.
authors.affiliation: Robotics and Biology Laboratory, Technische Universität Berlin, 10587, Berlin, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Schneider
authors.affiliation: Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh, EH9 3BF, United Kingdom.
authors.forename: Adam
authors.initials: A
authors.lastname: Belsom
authors.affiliation: Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh, EH9 3BF, United Kingdom. juri.rappsilber@ed.ac.uk.|Department of Bioanalytics, Institute of Biotechnology, Technische Universität Berlin, 13355, Berlin, Germany. juri.rappsilber@ed.ac.uk.
authors.forename: Juri
authors.initials: J
authors.lastname: Rappsilber
authors.affiliation: Robotics and Biology Laboratory, Technische Universität Berlin, 10587, Berlin, Germany. oliver.brock@tu-berlin.de.
authors.forename: Oliver
authors.initials: O
authors.lastname: Brock
mesh_terms: D000465:Algorithms; D000595:Amino Acid Sequence; D019295:Computational Biology / Q000379:methods / Q000706:statistics & numerical data*; D003198:Computer Simulation; D003432:Cross-Linking Reagents / Q000737:chemistry*; D030562:Databases, Protein; D007391:International Cooperation; D020407:Internet; D013058:Mass Spectrometry / Q000379:methods; D008958:Models, Molecular; D015233:Models, Statistical; D017510:Protein Folding; D054730:Protein Interaction Domains and Motifs; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry*; D016415:Sequence Alignment; D012984:Software; D013388:Succinimides / Q000737:chemistry*; D014466:Ultraviolet Rays
chemical_list: D003432:Cross-Linking Reagents; D011506:Proteins; D013388:Succinimides; C035760:bis(sulfosuccinimidyl)suberate
keywords: CASP; blind test; cross-linking; hybrid methods; mass spectrometry; protein structure prediction