Joint use of SAS and AUC

Olwyn Byron

Analytical ultracentrifugation (AUC) and small-angle x-ray and neutron scattering (SAXS, SANS or SAS overall) are fantastically complementary methods for characterising macromolecules in their near physiological, solution state [1, 2]. Each has its strengths and limitations, but used together, the combination is greater than the sum of its parts. In this lecture I will present the principles underlying AUC, describe how AUC experiments are typically performed, the types of systems that can be analysed and the data that result. I will focus on the interpretation of those data using hydrodynamic modelling (e.g. [3]), at which point I will stress the synergy between SAS and AUC (e.g. [4, 5, 6, 7]). The lecture will close by touching on new developments in hydrodynamic modelling of flexible systems and how these interface with SAS data and modelling.

1. Lebowitz, J., Lewis, M. S. & Schuck, P. (2002). Modern analytical ultracentrifugation in protein science: A tutorial review. Protein Science 11, 2067-2079.
2. Ralston, G. (1993). Introduction to Analytical Ultracentrifugation, Beckman Instruments, Inc., Palo Alto, California, USA.
3. Byron, O. (2008). Hydrodynamic modeling: The solution conformation of macromolecules and their complexes. In Methods in Cell Biology (Correia, J. J. & Detrich, H. W., eds.), Vol. 84, pp. 327-373. Elsevier.
4. Vijayakrishnan, S., Kelly, S. M., Gilbert, R. J. C., Callow, P., Bhella, D., Forsyth, T., Lindsay, J. G. & Byron, O. (2010). Solution structure and characterisation of the human pyruvate dehydrogenase complex core assembly. Journal of Molecular Biology 399, 71-93.
5. Vijayakrishnan, S., Callow, P., Nutley, M. A., McGow, D., Gilbert, D., Kropholler, P., Cooper, A., Byron, O. & Lindsay, J. G. (2011). Variation in the organisation and subunit composition of the mammalian pyruvate dehydrogenase complex E2/E3BP core assembly. Biochemical Journal 437, 565-574.
6. Gabrielsen, M., Beckham, K. S. H., Cogdell, R. J., Byron, O. & Roe, A. J. (2012). FolX from Pseudomonas aeruginosa is octameric in both crystal and solution. Febs Letters 586, 1160-1165.
7. Gabrielsen, M., Beckham, K. S. H., Feher, V. A., Zetterstrom, C. E., Wang, D., M?ller, S., Elofsson, M., Amaro, R. E., Byron, O. & Roe, A. J. (2012). Structural characterisation of Tpx from Yersinia pseudotuberculosis reveals insights into the binding of salicylidene acylhydrazide compounds. Plos One 7.

Date/time: Monday, 22 October 2012, 9:00