Preparation, crystallization and X-ray diffraction analysis to 1.5 Å resolution of rat cysteine dioxygenase, a mononuclear iron enzyme responsible for cysteine thiol oxidation

doi:10.1107/S1744309105033737

Title: Preparation, crystallization and X-ray diffraction analysis to 1.5 Å resolution of rat cysteine dioxygenase, a mononuclear iron enzyme responsible for cysteine thiol oxidation

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Abstract

Recombinant rat cysteine dioxygenase (CDO) has been expressed, purified and crystallized and X-ray diffraction data have been collected to 1.5 Å resolution. Cysteine dioxygenase (CDO; EC 1.13.11.20) is an ∼23 kDa non-heme iron metalloenzyme that is responsible for the oxidation of cysteine by O{sub 2}, yielding cysteinesulfinate. CDO catalyzes the first step in the conversion of cysteine to taurine, as well as the first step in the catabolism of cysteine to pyruvate plus sulfate. Recombinant rat CDO was heterologously expressed, purified and crystallized. The protein was expressed as a fusion protein bearing a polyhistidine tag to facilitate purification, a thioredoxin tag to improve solubility and a factor Xa cleavage site to permit removal of the entire N-terminus, leaving only the 200 amino acids inherent to the native protein. A multi-step purification scheme was used to achieve >95% purity of CDO. The optimal CDO crystals diffracted to 1.5 Å resolution and belonged to space group P4{sub 3}2{sub 1}2 or P4{sub 1}2{sub 1}2, with unit-cell parameters a = b = 57.55, c = 123.06 Å, α = β = γ = 90°. CDO shows little homology to any other proteins; therefore, the structure of the enzyme will be determined by ab initiomore »« less

Authors:

Simmons, Chad R. ^[1]; Hao, Quan ^[2]; Stipanuk, Martha H., E-mail: mhs6@cornell.edu ^[1]

Division of Nutritional Sciences, Cornell University, Ithaca, NY 14853-8001 (United States)
MacCHESS at the Cornell High Energy Synchrotron Source, Cornell University, Ithaca, NY 14853-8001 (United States)

Publication Date:: Tue Nov 01 00:00:00 EST 2005

OSTI Identifier:: 22356181

Resource Type:: Journal Article

Resource Relation:: Journal Name: Acta Crystallographica. Section F; Journal Volume: 61; Journal Issue: Pt 11; Other Information: PMCID: PMC1978133; PMID: 16511222; PUBLISHER-ID: gx5071; OAI: oai:pubmedcentral.nih.gov:1978133; Copyright (c) International Union of Crystallography 2005; Country of input: International Atomic Energy Agency (IAEA)

Country of Publication:: United Kingdom

Language:: English

Subject:: 75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CLEAVAGE; CONVERSION; CRYSTALS; CYSTEINE; HEME; IMPURITIES; IRON; OXIDATION; OXYGEN; RATS; RESOLUTION; SOLUBILITY; SPACE GROUPS; X-RAY DIFFRACTION

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                    Simmons, Chad R., Hao, Quan, and Stipanuk, Martha H., E-mail: mhs6@cornell.edu. Preparation, crystallization and X-ray diffraction analysis to 1.5 Å resolution of rat cysteine dioxygenase, a mononuclear iron enzyme responsible for cysteine thiol oxidation.  United Kingdom: N. p., 2005. 
        Web.  doi:10.1107/S1744309105033737.

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                    Simmons, Chad R., Hao, Quan, & Stipanuk, Martha H., E-mail: mhs6@cornell.edu. Preparation, crystallization and X-ray diffraction analysis to 1.5 Å resolution of rat cysteine dioxygenase, a mononuclear iron enzyme responsible for cysteine thiol oxidation.  United Kingdom.  doi:10.1107/S1744309105033737.

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                    Simmons, Chad R., Hao, Quan, and Stipanuk, Martha H., E-mail: mhs6@cornell.edu. Tue .  
        "Preparation, crystallization and X-ray diffraction analysis to 1.5 Å resolution of rat cysteine dioxygenase, a mononuclear iron enzyme responsible for cysteine thiol oxidation".  United Kingdom.  
        doi:10.1107/S1744309105033737.

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@article{osti_22356181,

  title        = {Preparation, crystallization and X-ray diffraction analysis to 1.5 Å resolution of rat cysteine dioxygenase, a mononuclear iron enzyme responsible for cysteine thiol oxidation},

  author       = {Simmons, Chad R. and Hao, Quan and Stipanuk, Martha H., E-mail: mhs6@cornell.edu},

  abstractNote = {Recombinant rat cysteine dioxygenase (CDO) has been expressed, purified and crystallized and X-ray diffraction data have been collected to 1.5 Å resolution. Cysteine dioxygenase (CDO; EC 1.13.11.20) is an ∼23 kDa non-heme iron metalloenzyme that is responsible for the oxidation of cysteine by O{sub 2}, yielding cysteinesulfinate. CDO catalyzes the first step in the conversion of cysteine to taurine, as well as the first step in the catabolism of cysteine to pyruvate plus sulfate. Recombinant rat CDO was heterologously expressed, purified and crystallized. The protein was expressed as a fusion protein bearing a polyhistidine tag to facilitate purification, a thioredoxin tag to improve solubility and a factor Xa cleavage site to permit removal of the entire N-terminus, leaving only the 200 amino acids inherent to the native protein. A multi-step purification scheme was used to achieve >95% purity of CDO. The optimal CDO crystals diffracted to 1.5 Å resolution and belonged to space group P4{sub 3}2{sub 1}2 or P4{sub 1}2{sub 1}2, with unit-cell parameters a = b = 57.55, c = 123.06 Å, α = β = γ = 90°. CDO shows little homology to any other proteins; therefore, the structure of the enzyme will be determined by ab initio phasing using a selenomethionyl derivative.},

  doi          = {10.1107/S1744309105033737},

  journal      = {Acta Crystallographica. Section F},

  number       = Pt 11,

  volume       = 61,

  place        = {United Kingdom},

  year         = {Tue Nov 01 00:00:00 EST 2005},

  month        = {Tue Nov 01 00:00:00 EST 2005}

}

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DOI: 10.1107/S1744309105033737

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Preparation, Crystallization and X-ray Diffraction Analysis to 1.5 A Resolution of Rat Cysteine Dioxygenase, a Mononuclear Iron Enzyme Responsible for Cysteine Thiol Oxidation

Simmons,C. ; Hao, Q. ; Stipanuk, M. - Acta Cryst. F

Cysteine dioxygenase (CDO; EC 1.13.11.20) is an {approx}23 kDa non-heme iron metalloenzyme that is responsible for the oxidation of cysteine by O2, yielding cysteinesulfinate. CDO catalyzes the first step in the conversion of cysteine to taurine, as well as the first step in the catabolism of cysteine to pyruvate plus sulfate. Recombinant rat CDO was heterologously expressed, purified and crystallized. The protein was expressed as a fusion protein bearing a polyhistidine tag to facilitate purification, a thioredoxin tag to improve solubility and a factor Xa cleavage site to permit removal of the entire N-terminus, leaving only the 200 amino acidsmore »« less
DOI: 10.1107/S1744309105033737
Crystal Structure of Mammalian Cysteine dioxygenase: A Novel Mononuclear Iron Center for Cysteine Thiol Oxidation

Simmons,C. ; Liu, Q. ; Huang, Q. ; ... - J. Biol. Chem.

Cysteine dioxygenase is a mononuclear iron-dependent enzyme responsible for the oxidation of cysteine with molecular oxygen to form cysteinesulfinate. This reaction commits cysteine to either catabolism to sulfate and pyruvate or to the taurine biosynthetic pathway. Cysteine dioxygenase is a member of the cupin superfamily of proteins. The crystal structure of recombinant rat cysteine dioxygenase has been determined to 1.5 Angstroms resolution, and these results confirm the canonical cupin {beta}-sandwich fold and the rare cysteinyl-tyrosine intramolecular crosslink (between Cys93 and Tyr157) seen in the recently reported murine cysteine dioxygenase structure. In contrast to the catalytically inactive mononuclear Ni(II) metallocenter presentmore »« less
DOI: 10.1074/jbc.M601555200
Crystallization and preliminary X-ray diffraction analysis of the electron-transfer complex between the terminal oxygenase component and ferredoxin in the Rieske non-haem iron oxygenase system carbazole 1,9a-dioxygenase

Ashikawa, Yuji ; Fujimoto, Zui ; Noguchi, Haruko ; ... - Acta Crystallographica. Section F

The electron-transfer complex between the terminal oxygenase and ferredoxin of carbazole 1,9a-dioxygenase was crystallized and diffraction data were collected to 1.90 Å resolution. Carbazole 1,9a-dioxygenase, which consists of an oxygenase component (CARDO-O) and the electron-transport components ferredoxin (CARDO-F) and ferredoxin reductase (CARDO-R), catalyzes dihydroxylation at the C1 and C9a positions of carbazole. The electron-transport complex between CARDO-O and CARDO-F crystallizes at 293 K using hanging-drop vapour diffusion with the precipitant PEG MME 2000 (type I crystals) or PEG 3350 (type II). Blossom-shaped crystals form from a pile of triangular plate-shaped crystals. The type I crystal diffracts to a maximum resolutionmore »« less
DOI: 10.1107/S1744309105014557
Human tRNA{sup Gly} acceptor-stem microhelix: crystallization and preliminary X-ray diffraction analysis at 1.2 Å resolution

Förster, Charlotte ; Szkaradkiewicz, Karol ; Perbandt, Markus ; ... - Acta Crystallographica. Section F

The human tRNA{sup Gly} acceptor-stem microhelix was crystallized and preliminary X-ray diffraction analysis revealed diffraction to a resolution of up to 1.2 Å. The major dissimilarities between the eukaryotic/archaebacterial-type and eubacterial-type glycyl-tRNA synthetase systems (GlyRS; class II aminoacyl-tRNA synthetases) represent an intriguing example of evolutionarily divergent solutions to similar biological functions. The differences in the identity elements of the respective tRNA{sup Gly} systems are located within the acceptor stem and include the discriminator base U73. In the present work, the human tRNA{sup Gly} acceptor-stem microhelix was crystallized in an attempt to analyze the structural features that govern the correct recognitionmore »« less
DOI: 10.1107/S1744309107041528
Cloning, expression, purification, crystallization and X-ray diffraction analysis of dihydrodipicolinate synthase from the human pathogenic bacterium Bartonella henselae strain Houston-1 at 2.1 Å resolution

Naqvi, Kubra F. ; Staker, Bart L. ; Dobson, Renwick C. J. ; ... - Acta Crystallographica. Section F, Structural Biology Communications

The enzyme dihydrodipicolinate synthase catalyzes the committed step in the synthesis of diaminopimelate and lysine to facilitate peptidoglycan and protein synthesis. Dihydrodipicolinate synthase catalyzes the condensation of L-aspartate 4-semialdehyde and pyruvate to synthesize L-2,3-dihydrodipicolinate. Here, the cloning, expression, purification, crystallization and X-ray diffraction analysis of dihydrodipicolinate synthase from the pathogenic bacteriumBartonella henselae, the causative bacterium of cat-scratch disease, are presented. Protein crystals were grown in conditions consisting of 20%(w/v) PEG 4000, 100 mMsodium citrate tribasic pH 5.5 and were shown to diffract to ~2.10 Å resolution. They belonged to space groupP2 ₁2 ₁2 ₁, with unit-cell parametersa= 79.96,b= 106.33,c= 136.25more »« less
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