title: Conformational spectra--probing protein conformational changes.
abstract: Stafford [Biophys. J. 17 (1996) MP452] has shown that it is possible, using the analytical ultracentrifuge in sedimentation velocity mode, to calculate the molecular weights of proteins with a precision of approximately 5%, by fitting Gaussian distributions to g(s*) profiles so long as partial specific volume and the radial position of the meniscus are known. This makes possible the analysis of systems containing several components by the fitting of multiple distributions to the total g(s*) profile. We have found the Stafford relationship to hold for a range of protein solutes, particularly good agreement being found when the g(s*) profiles are computed from Schlieren (dc/dr vs. r) data using the Bridgman equation [J. Am. Chem. Soc. 64 (1942) 2349] . On this basis, we have developed a new approach to the analysis of systems where two or more distinguishable conformations of a single species are present, either in the same sample cell or in different cells in the same rotor. In the former case, this allows us to analyse a given solution of pure protein (i.e. monodisperse with respect to M) to reveal the presence in that solution of two or more conformers under identical solvent conditions. In the latter case, we can detect with high sensitivity any conformational change occurring in the transition from one set of solvent conditions to another. Alternatively, in this case, we can analyse slightly different proteins (e.g. deletion mutants) for conformational changes under identical solvent conditions. Examples of these procedures using well-defined protein systems are given.
authors.affiliation: NCMH Business Centre, School of Biological Sciences, University of Nottingham, Sutton Bonington, Leicestershire LE12 5RD, UK. neil.errington@nottingham.ac.uk
authors.forename: N
authors.initials: N
authors.lastname: Errington
authors.forename: O
authors.initials: O
authors.lastname: Byron
authors.forename: A J
authors.initials: AJ
authors.lastname: Rowe