International
Tables for Crystallography Volume F Crystallography of biological macromolecules Edited by M. G. Rossmann and E. Arnold © International Union of Crystallography 2006 |
International Tables for Crystallography (2006). Vol. F. ch. 19.4, pp. 442-443
Section 19.4.5.1. Feasibility
aDepartment of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520, USA, and bDepartments of Chemistry and Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520, USA |
For a biological scientist, the first question is whether knowledge of the parameters that can be obtained from neutron scattering is of interest for the biological problem under consideration. If the answer is affirmative, the best course of action is to make contact with individuals who have conducted experiments in the past, as well as with biologists at a neutron-scattering facility. As a preliminary guide, a few general points are pertinent.
For a solution-scattering experiment on an unlabelled sample, typical sample volumes are 0.1–0.2 ml at concentrations of around 10 mg ml−1. For contrast variation, samples dialysed to different D2O levels will be required, so significant amounts of material need to be at hand. Making a set of measurements will require a few hours at a modern facility.
Longer collection times and significantly greater biochemical efforts will be involved for measurements with labelled material, especially if reconstitution is part of the strategy. Labelled biomolecules are most often produced by growth of organisms in D2O (Moore & Engelman, 1976; Vanatalu et al., 1993
), but strategies using chemical synthesis or providing labelled precursors have also been employed. Preparation of samples with labelled ligands is usually more straightforward than the generation of reconstituted complexes, but still requires tests of homogeneity.
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