TOP

Research Article

Split Viewer

Mol. Cells 2013; 36(1): 74-81

Published online May 16, 2013

https://doi.org/10.1007/s10059-013-0074-1

© The Korean Society for Molecular and Cellular Biology

In-House Zinc SAD Phasing at Cu Kα Edge

Min-Kyu Kim, Sangmin Lee, Young Jun An, Chang-Sook Jeong, Chang-Jun Ji, Jin-Won Lee, and Sun-Shin Cha

1Marine Biotechnology Research Division, Korea Institute of Ocean Science and Technology, Ansan 426-744, Korea, 2Ocean Science and Technology School, Pusan 606-791, Korea, 3Department of Life Science and Institute for Natural Sciences, Hanyang University, Seoul 133-791, Korea, 4Department of Marine Biotechnology, University of Science and Technology, Daejeon 305-333 Korea, 5These authors contributed equally to this work.

Received: February 28, 2013; Revised: April 9, 2013; Accepted: April 11, 2013

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 Kα 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.

Keywords anomalous scattering, experimental phasing, protein crystallography, SAD phasing, zinc

Article

Research Article

Mol. Cells 2013; 36(1): 74-81

Published online July 31, 2013 https://doi.org/10.1007/s10059-013-0074-1

Copyright © The Korean Society for Molecular and Cellular Biology.

In-House Zinc SAD Phasing at Cu Kα Edge

Min-Kyu Kim, Sangmin Lee, Young Jun An, Chang-Sook Jeong, Chang-Jun Ji, Jin-Won Lee, and Sun-Shin Cha

1Marine Biotechnology Research Division, Korea Institute of Ocean Science and Technology, Ansan 426-744, Korea, 2Ocean Science and Technology School, Pusan 606-791, Korea, 3Department of Life Science and Institute for Natural Sciences, Hanyang University, Seoul 133-791, Korea, 4Department of Marine Biotechnology, University of Science and Technology, Daejeon 305-333 Korea, 5These authors contributed equally to this work.

Received: February 28, 2013; Revised: April 9, 2013; Accepted: April 11, 2013

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 Kα 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.

Keywords: anomalous scattering, experimental phasing, protein crystallography, SAD phasing, zinc

Mol. Cells
Nov 30, 2023 Vol.46 No.11, pp. 655~725
COVER PICTURE
Kim et al. (pp. 710-724) demonstrated that a pathogen-derived Ralstonia pseudosolanacearum type III effector RipL delays flowering time and enhances susceptibility to bacterial infection in Arabidopsis thaliana. Shown is the RipL-expressing Arabidopsis plant, which displays general dampening of the transcriptional program during pathogen infection, grown in long-day conditions.

Share this article on

  • line

Related articles in Mol. Cells

Molecules and Cells

eISSN 0219-1032
qr-code Download