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Title: Crystallization, data collection and phasing of the molybdate-binding protein of the phytopathogen Xanthomonas axonopodis pv. citri

Abstract

The molybdate-binding protein (ModA) from X. axonopodis pv. citri was crystallized with sodium molybdate in the presence of PEG or sulfate. The crystal diffracted to a maximum resolution of 1.7 Å and belongs to the orthorhombic space group C222{sub 1,} with unit-cell parameters a = 68.15, b = 172.14, c = 112.04 Å. Xanthomonas axonopodis pv. citri ModA protein is the ABC periplasmic binding component responsible for the capture of molybdate. The protein was crystallized with sodium molybdate using the hanging-drop vapour-diffusion method in the presence of PEG or sulfate. X-ray diffraction data were collected to a maximum resolution of 1.7 Å using synchrotron radiation. The crystal belongs to the orthorhombic space group C222{sub 1}, with unit-cell parameters a = 68.15, b = 172.14, c = 112.04 Å. The crystal structure was solved by molecular-replacement methods and structure refinement is in progress.

Authors:
; ;  [1];  [2];  [3]
  1. Departamento de Microbiologia, Instituto de Ciências Biomédicas II, Universidade de São Paulo, São Paulo, SP (Brazil)
  2. Centro de Biologia Molecular e Estrutural (CeBiMe), Laboratório Nacional de Luz Síncrotron (LNLS), CP 6192, Campinas, SP 13084-971 (Brazil)
  3. (Brazil)
Publication Date:
OSTI Identifier:
22356297
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 62; Journal Issue: Pt 3; Other Information: PMCID: PMC2197186; PMID: 16511325; PUBLISHER-ID: ll5045; OAI: oai:pubmedcentral.nih.gov:2197186; Copyright (c) International Union of Crystallography 2006; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CAPTURE; CRYSTAL STRUCTURE; CRYSTALLIZATION; CRYSTALS; DIFFUSION; MOLYBDATES; PROTEINS; RESOLUTION; SODIUM; SPACE GROUPS; SULFATES; SYNCHROTRON RADIATION; X-RAY DIFFRACTION

Citation Formats

Santacruz, C. P., Balan, A., Ferreira, L. C. S., Barbosa, J. A. R. G., E-mail: joao@lnls.br, and Departamento de Microbiologia, Instituto de Ciências Biomédicas II, Universidade de São Paulo, São Paulo, SP. Crystallization, data collection and phasing of the molybdate-binding protein of the phytopathogen Xanthomonas axonopodis pv. citri. United Kingdom: N. p., 2006. Web. doi:10.1107/S1744309106003812.
Santacruz, C. P., Balan, A., Ferreira, L. C. S., Barbosa, J. A. R. G., E-mail: joao@lnls.br, & Departamento de Microbiologia, Instituto de Ciências Biomédicas II, Universidade de São Paulo, São Paulo, SP. Crystallization, data collection and phasing of the molybdate-binding protein of the phytopathogen Xanthomonas axonopodis pv. citri. United Kingdom. doi:10.1107/S1744309106003812.
Santacruz, C. P., Balan, A., Ferreira, L. C. S., Barbosa, J. A. R. G., E-mail: joao@lnls.br, and Departamento de Microbiologia, Instituto de Ciências Biomédicas II, Universidade de São Paulo, São Paulo, SP. Wed . "Crystallization, data collection and phasing of the molybdate-binding protein of the phytopathogen Xanthomonas axonopodis pv. citri". United Kingdom. doi:10.1107/S1744309106003812.
@article{osti_22356297,
title = {Crystallization, data collection and phasing of the molybdate-binding protein of the phytopathogen Xanthomonas axonopodis pv. citri},
author = {Santacruz, C. P. and Balan, A. and Ferreira, L. C. S. and Barbosa, J. A. R. G., E-mail: joao@lnls.br and Departamento de Microbiologia, Instituto de Ciências Biomédicas II, Universidade de São Paulo, São Paulo, SP},
abstractNote = {The molybdate-binding protein (ModA) from X. axonopodis pv. citri was crystallized with sodium molybdate in the presence of PEG or sulfate. The crystal diffracted to a maximum resolution of 1.7 Å and belongs to the orthorhombic space group C222{sub 1,} with unit-cell parameters a = 68.15, b = 172.14, c = 112.04 Å. Xanthomonas axonopodis pv. citri ModA protein is the ABC periplasmic binding component responsible for the capture of molybdate. The protein was crystallized with sodium molybdate using the hanging-drop vapour-diffusion method in the presence of PEG or sulfate. X-ray diffraction data were collected to a maximum resolution of 1.7 Å using synchrotron radiation. The crystal belongs to the orthorhombic space group C222{sub 1}, with unit-cell parameters a = 68.15, b = 172.14, c = 112.04 Å. The crystal structure was solved by molecular-replacement methods and structure refinement is in progress.},
doi = {10.1107/S1744309106003812},
journal = {Acta Crystallographica. Section F},
number = Pt 3,
volume = 62,
place = {United Kingdom},
year = {Wed Mar 01 00:00:00 EST 2006},
month = {Wed Mar 01 00:00:00 EST 2006}
}
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  • Cloning, expression, purification, crystallization and data collection are reported for a member of the SufE family of proteins involved in the biosynthesis of Fe–S clusters in prokaryotes. Diffraction data were collected to 1.9 Å resolution and an interpretable electron-density map has been obtained by molecular replacement. Xanthomonas axonopodis pv. citri (Xac) SufE (XAC2355) is a member of a family of bacterial proteins that are conserved in several pathogens and phytopathogens. The Escherichia coli suf operon is involved in iron–sulfur cluster biosynthesis under iron-limitation and stress conditions. It has recently been demonstrated that SufE and SufS form a novel two-component cysteinemore » desulfarase in which SufS catalyses the conversion of l-cysteine to l-alanine, forming a protein-bound persulfide intermediate. The S atom is then transferred to SufE, from which it is subsequently transferred to target molecules or reduced to sulfide in solution. Here, the cloning, expression, crystallization and phase determination of Xac SufE crystals are described. Recombinant SufE was crystallized in space group P2{sub 1}2{sub 1}2{sub 1} and diffracted to 1.9 Å resolution at a synchrotron source. The unit-cell parameters are a = 45.837, b = 58.507, c = 98.951 Å, α = β = γ = 90°. The calculated Matthews coefficient indicated the presence of two molecules in the asymmetric unit. Phasing was performed by molecular-replacement using E. coli SufE as a model (PDB code 1mzg) and an interpretable map was obtained.« less
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