Biological small angle X-ray scattering (bioSAXS) is a technique for obtaining ‘low’ resolution (~nm) structural information from macromolecules in solution that is complementary to Macromolecular Crystallography (MX) and becoming more widely used in structural biology. The bioSAXS beamline on BM29 at the ESRF has an optimised optical layout for solution scattering experiments with a multilayer monochromator for maximising the available flux and a toroidal focusing mirror for the generation of a small 100 µm2 beamsize. The beamline is currently equipped with a PILATUS 1M detector and a liquid handling robot designed by the Diffraction Instrumentation Team as part of a trilateral agreement between the ESRF and both the EMBL Grenoble and Hamburg outstations. A Shimatzu HPLC system with an auto-sampler is also available for optimal experimental SAXS measurements from difficult biological systems.
Adam Round of the Synchrotron Crystallography Team leads the EMBL Grenoble contribution to this joint effort. Currently, users can investigate a wide range of proteins under physiological conditions and undertake functional studies investigating the effects of differing temperatures (2-50 degrees oC), pH (4-10), salt concentration and/or the addition of ligands in samples as small as 5 µl (25-30 µl are recommended). The bioSAXS beamline is dedicated to measuring biological macromolecules in solution with the aim of providing a simple to use facility with rapid access using the ESRF MX BAG or rolling application procedures. A high level of automation is implemented to aid users with data collection, rapid preliminary analysis and online quality control using an EDNA implementation of the data analysis tools developed by EMBL Hamburg and displayed in ISPyB for bioSAXS (ISPyBB). This provides users with valuable feedback regarding their experiment to maximise throughput, dataset completeness and the possible need for additional measurements.
As part of the PSB SAS platform there is a unified application procedure within the ILL application procedure to enable joint SAXS/SANS experiments at the ILL and the ESRF facilities in one visit to the EPN site.
The BioSAXS beamline is run as a collaboration between the EMBL Grenoble Synchrotron Crystallography Team and the ESRF Structural Biology Group. See the beamline webpage for more information on the technical details and the ESRF website for application details.
De Maria Antolinos, A., Pernot, P., Brennich, M. E., Kieffer, J., Bowler, M. W., Delageniere, S., Ohlsson, S., Malbet Monaco, S., Ashton, A., Franke, D., Svergun, D., McSweeney, S., Gordon, E., Round, A. (2015) ISPYB for BioSAXS, the gateway to user autonomy in solution scattering experiments. Acta Crystallogr. D Biol. Crystallogr. 71, 76-85.
Round, A., Felisaz, F., Fodinger, L., Gobbo, A., Huet, J., Villard, C., Blanchet, C. E., Pernot, P. McSweeney, S., Roessle, M., Svergun, D. I., Cipriani, F. (2015) BioSAXS Sample Changer: a robotic sample changer for rapid and reliable high-throughput X-ray solution scattering experiments (2015) Acta Crystallogr. D Biol. Crystallogr. 71, 67-75.
Pernot, P., Round, A., Barrett, R., De Maria Antolinos, A., Gobbo, A., Gordon, E., Huet, J., Kieffer, J., Lentini, M., Mattenet, M., Morawe, C., Mueller-Dieckmann, C., Ohlsson, S., Schmid, W., Surr, J., Theveneau, P., Zerrad, L., McSweeney, S. (2013) Upgraded ESRF BM29 beamline for SAXS on macromolecules in solution. J. Synchrotron Rad. 20, 660-664.
BM29 research highlights in 2015-2016
Thierry, E., Guilligay, D., Kosinski, J., Bock, T., Gaudon, S., Round, A., Pflug, A., Hengrung, N., El Omari, K., Baudin, F., Hart, D. J., Beck, M., Cusack, S. (2016) Influenza Polymerase Can Adopt an Alternative Configuration Involving a Radical Repacking of PB2 Domains. Mol. Cell
Delaforge, E., Milles, S., Bouvignies, G., Bouvier, D., Boivin, S., Salvi, N., Maurin, D., Martel, A., Round, A., Lemke, E. A., Ringkjøbing Jensen, M., Hart, D. J., Blackledge, M. (2015) J. Am. Chem. Soc. 137, 15122-15134.