In-house zinc SAD phasing at Cu K alpha edge

Abstract

De novo zinc single-wavelength anomalous dispersion (Zn-SAD) phasing has been demonstrated with the 1.9 resolution data of glucose isomerase and 2.6 resolution data of Staphylococcus aureus Fur (SaFur) collected using in-house Cu K alpha X-ray source. The successful in-house Zn-SAD phasing of glucose isomerase, based on the anomalous signals of both zinc ions introduced to crystals by soaking and native sulfur atoms, drove us to determine the structure of SaFur, a zinc-containing transcription factor, by Zn-SAD phasing using in-house X-ray source. The abundance of zinc-containing proteins in nature, the easy zinc derivatization of the protein surface, no need of synchrotron access, and the successful experimental phasing with the modest 2.6 resolution SAD data indicate that inhouse Zn-SAD phasing can be widely applicable to structure determination.