Journal:Acta Cryst F:S2053230X19004151
From Proteopedia
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<b>Molecular Tour</b><br> | <b>Molecular Tour</b><br> | ||
| - | With better tools for data processing and synchrotron beamlines capable of collecting data at longer wavelengths, sulfur-based native single-wavelength anomalous dispersion (SAD) phasing has become the "first-choice" method for ''de novo'' protein structure determination. However, for many proteins, native SAD phasing can be simplified by taking advantage of their interactions with natural metal cofactors that are stronger anomalous scatterers than sulfur. This is demonstrated here for four unique domains of a 1.5-megadalton calcium-dependent adhesion protein using the anomalous diffraction of the chelated calcium ions. In all cases, low anomalous multiplicity X-ray data were collected on a home-source diffractometer equipped with a chromium rotating anode (λ = 2.2909 Å). In all but one case, calcium-SAD phasing alone was sufficient to allow automated model building and refinement of the protein model after the calcium substructure was determined. Given that calcium atoms will be present in a significant percentage of the proteins that remain uncharacterized, many aspects of the data collection and processing methods described here could be broadly applied for routine ''de novo'' structure elucidation. | + | With better tools for data processing and synchrotron beamlines capable of collecting data at longer wavelengths, sulfur-based native single-wavelength anomalous dispersion (SAD) phasing has become the "first-choice" method for ''de novo'' protein structure determination. However, for many proteins, native SAD phasing can be simplified by taking advantage of their interactions with natural metal cofactors that are stronger anomalous scatterers than sulfur. This is demonstrated here for four unique domains of a 1.5-megadalton calcium-dependent adhesion protein using the anomalous diffraction of the chelated calcium ions. In all cases, low anomalous multiplicity X-ray data were collected on a home-source diffractometer equipped with a chromium rotating anode (λ = 2.2909 Å). In all but one case, calcium-SAD phasing alone was sufficient to allow automated model building and refinement of the protein model after the calcium substructure was determined. Given that calcium atoms will be present in a significant percentage of the proteins that remain uncharacterized, many aspects of the data collection and processing methods described here could be broadly applied for routine ''de novo'' structure elucidation. |
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| + | X-ray crystal structures solved by Ca-SAD phasing: | ||
| + | *<scene name='81/812834/Cv/2'>TextToBeDisplayed</scene> RII-monomer (PDB [[4kdv]]; cyan). | ||
<b>References</b><br> | <b>References</b><br> | ||
Revision as of 09:14, 14 April 2019
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This page complements a publication in scientific journals and is one of the Proteopedia's Interactive 3D Complement pages. For aditional details please see I3DC.
