Abstract
By varying the relative values of protein and solvent scattering densities in a crystal, it is possible to obtain information on the shape and dimensions of protein molecular envelopes. Neutron diffraction methods are ideally suited to these contrast variation experiments because H/D exchange leads to large differential changes in the protein and solvent scattering densities and is structurally non-perturbing. Low resolution structure factors have been measured from cubic insulin crystals with differing H/D contents. Structure factors calculated from a simple binary density model, in which uniform scattering densities represent the protein and solvent volumes in the crystals, were compared with these data. The contrast variation differences in the sets of measured structure factors were found to be accurately fitted by this simple model. Trial applications to two problems in crystal structure determination illustrate how this fact may be exploited, (i) A translation function that employs contrast variation data gave a sharp minimum within 1–9Å of the correctly positioned insulin molecule and is relatively insensitive to errors in the atomic model, (ii) An ab initio phasing method for the contrast variation data, based on analyzing histograms of the density distributions in trial maps, was found to recover the correct molecular envelope.
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Badger, J. (1996). Determination of Protein and Solvent Volumes in Protein Crystals from Contrast Variation Data. In: Schoenborn, B.P., Knott, R.B. (eds) Neutrons in Biology. Basic Life Sciences, vol 64. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-5847-7_29
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DOI: https://doi.org/10.1007/978-1-4615-5847-7_29
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