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• W2156875598 abstract "The basic theory and principles of the multiple-wavelength anomalous solvent-contrast (MASC) method are introduced as a contrast-variation technique for generating low-resolution crystallographic phase information on the envelope of a macromolecule. Experimental techniques and practical considerations concerning the choice of anomalous scatterer, sample preparation and data acquisition are discussed. Test cases of crystals of three proteins of differing molecular weights from 14 kDa through to 173 kDa are illustrated. Methods for extracting the moduli of the anomalous structure factors from the MASC data are briefly discussed and the experimental results are compared with the known macromolecular envelopes. In all cases, the lowest resolution shells exhibit very large anomalous signals which diminish at higher resolution, as expected by theory. However, in each case the anomalous signal persists at high resolution, which is strong evidence for ordered sites of the anomalous scatterers. For the smaller two of these proteins the heavy-atom parameters could be refined for some of these sites. Finally, a novel method for phasing the envelope structure-factor moduli is presented. This method takes into account the relatively low number of observations at low resolution and describes the macromolecular envelope with a small number of parameters by presuming that the envelope is a compact domain of known volume. The parameterized envelope is expressed as a linear combination of independent functions such as spherical harmonics. Phasing starts from solutions of a sphere in the unit cell after positional refinement from random trials and the parameters describing the envelope are then refined against the data of structure-factor moduli. The preliminary results using simulated data show that the method can be used to reconstruct the correct macromolecular envelope and is able to discriminate against some false solutions." @default.
• W2156875598 created "2016-06-24" @default.
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• W2156875598 date "2000-10-01" @default.
• W2156875598 modified "2023-09-26" @default.
• W2156875598 title "Low-resolution phase information in multiple-wavelength anomalous solvent contrast variation experiments" @default.
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• W2156875598 doi "https://doi.org/10.1107/s0907444900009574" @default.
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