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Lookup NU author(s): Dr Paula SalgadoORCiD
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SAD phasing has been revisited recently, with experiments being carried out using previously unconventional sources of anomalous signal, particularly lighter atoms and softer X-rays. A case study is reported using the 75 kDa RNA-dependent RNA polymerase of the bacteriophase [varphi]6, which binds a Mn atom and crystallizes with three molecules in the asymmetric unit. X-ray diffraction data were collected at a wavelength of 1.89 Å and although the calculated anomalous signal from the three Mn atoms was only 1.2%, SHELXD and SOLVE were able to locate these atoms. SOLVE/RESOLVE used this information to obtain SAD phases and automatically build a model for the core region of the protein, which possessed the characteristic features of the right-hand polymerase motif. These results demonstrate that with modern synchrotron beamlines and software, manganese phasing is a practical tool for solving the structure of large proteins.
Author(s): Salgado PS, Walsh MA, Laurila MRL, Stuart DI, Grimes JM
Publication type: Article
Publication status: Published
Journal: Acta Biol Crystallographica D
ISSN (print): 0907-4449
ISSN (electronic): 1399-0047
Publisher: Wiley-Blackwell Publishing, Inc.
PubMed id: 15608382
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