This EMBO Practical Course aims to teach participants how to expedite structural biology projects involving macromolecular complexes by combining multiple experimental approaches. In particular, the course will cover the techniques used to produce, purify, reconstitute and characterize multi-subunit protein and protein/nucleic acid complexes for structural analysis. This course is primarily aimed at PhD students and postdocs working on challenging structural projects involving macromolecular complexes.
Topics include: multi-subunit expression in bacteria, insect and mammalian cells; affinity, electrophoretic and centrifugation methods to isolate and purify sub-complexes and holo-complexes; biochemical and biophysical experiments that inform on the stoichiometric and conformational homogeneity of complexes; and strategies for determining the overall 3D architecture of large macromolecular assemblies and ultimately their structure at atomic or pseudo-atomic resolution.
Special emphasis will be placed on: (i) the latest advances in isolating complexes from endogenous sources, in recombinantly co-expressing binary and higher-order combinations of complex components, and in reconstituting complexes from individually purified components or sub-complexes;(ii) techniques for assessing and improving sample quality (homogeneity, stoichiometry, monodispersity); (iii) combining structure determination methods (crystallography, cryo-electron microscopy, small-angle scattering, NMR , native mass spectrometry, etc.) to obtain a multi- resolution view of a given complex.
Speakers: Radu Aricescu (University of Oxford), Imre Berger (University of Bristol), Elisabetta Boeri Erba (Institut de Biologie Structurale), Toby Gibson (EMBL Heidelberg), Gordon Leonard (European Synchrotron Radiation Facility), Joe Marsh (University of Edinburgh), Adrien Melquiond (Utrecht University), Sigrid Milles (Institut de Biologie Structurale), Robert Rambo (Diamond Light Source), Jonas Ries (EMBL Heidelberg), Malene Jensen Ringkjobing (Institut de Biologie Structurale), Christophe Romier (Institut de Génétique et de Biologie Moléculaire et Cellulaire) , Vladimir Rybin (EMBL Heidelberg), Helen Saibil (Birkbeck College), Michael Sattler (Technische Universität München), Bertrand Séraphin (Institut de Génétique et de Biologie Moléculaire et Cellulaire), Andrea Sinz (Martin Luther University of Halle-Wittenberg), Holger Stark (Max Planck Institute), Jan Steyaert (Vrije Universiteit Brussel), Song Tan (Penn State University), Navtej Toor (University of California San Diego), Jill Trewhella (University of Sydney), Christine Ziegler (University of Regensburg)
Travel grants: A limited number of travel grants are available for eligible participants. (Applicants do not need to apply separately for travel grants. Selection of awardees is handled directly by the organisers who will notify all eligible participants).
Course Organizers: Carlo Petosa (Institut de Biologie Structurale), Trevor Forsyth (Institut Laue-Langevin FR and Keele University), Darren Hart (Institut de Biologie Structurale), Montserrat Soler López (European Synchrotron Radiation Facility), Marco Marcia (EMBL Grenoble), Daniele de Sanctis (European Synchrotron Radiation Facility)