1.13.11.20: cysteine dioxygenase

This is an abbreviated version!
For detailed information about cysteine dioxygenase, go to the full flat file.

Word Map on EC 1.13.11.20

Reaction

L-cysteine
+
O2
=
3-sulfinoalanine

Synonyms

3-mercaptopropionate dioxygenase, Arg-type CDO, BsCDO, CDO, CDO1, CDO2, CdoA, CdoB, cysteine dioxygenase, cysteine oxidase, Fe(II) cysteine dioxygenase, H16_A1614, H16_B1863, oxygenase, cysteine di-

ECTree

     1 Oxidoreductases
         1.13 Acting on single donors with incorporation of molecular oxygen (oxygenases)
             1.13.11 With incorporation of two atoms of oxygen
                1.13.11.20 cysteine dioxygenase

Purification

Purification on EC 1.13.11.20 - cysteine dioxygenase

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PURIFICATION/commentary
ORGANISM
UNIPROT
LITERATURE
1 ml HisTrap HP column, metal ion (nickel) affinity chromatography, gel filtration, MonoQ 4.6/100 PE ion exchange column. The thioredoxin/6x His cysteine dioxygenase fusion protein separates into two apparent isoforms that elute as two distinct peaks, one that elutes at 50 mM imidazole and one that elutes at 100 mM imidazole during metal ion affinity chromatograohy. The protein in the second peak has a specific activity, that is 50-60% less than that of the protein in the first peak. In contrast to peak 1 peak 2 elutes as two pronounced peaks (A and B) from the MonoQ column. The cysteine dioxygenase in peak A has no detectabel activity. In the the standard cysteine dioxygenase purification procedure, only the form from peak 1 is retained and further purified.
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all buffers are supplemented with 5 mM dithiothreitol to prevent oxidation of the selenomethionine
amylose column chromatography and Superdex 200 gel filtration
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HisTrap HP column, immobilized metal affinity chromatography, during purification two peaks of highly purified recombinant protein are found to elute: one at 10% (peak1) and the second at 20% (peak 2), kinetic data show that peak 1 has lower Km and higher Vmax values for cysteine than peak 2
HisTrap HP column, immobilized metal affinity chromatography, during purification two peaks of highly purified recombinant protein are found to elute: one at 10% (peak1) and the second at 20%(peak 2), kinetic datat show that peak 1 has lower Km and higher Vmax values for cysteine thant peak 2.
HisTrap HP column, immobilized metal affinity chromatography; HisTrap HP column, immobilized metal affinity chromatography, during purification of SCO30335 two peaks of highly purified recombinant protein are found to elute: one at 10% (peak1) and the second at 20% (peak 2), kinetic data show that peak 1 has lower Km and higher Vmax values for cysteine than peak 2
O50490, Q9KZL0
immobilized metal chelate affinity chromatography
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immobilized nickel affinity chromatography
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Ni-NTA agarose column chromatography
recombinant enzyme
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recombinant enzyme from Escherichia coli strain BL21(DE3)
recombinant His-tagged enzyme CdoA from Escherichia coli by nickel affinity chromatography to homogeneity; recombinant His-tagged enzyme CdoB from Escherichia coli by nickel affinity chromatography to homogeneity
recombinant N-terminal maltose-binding protein fusion enzyme from Escherichia coli strain BL21(DE3) by anion exchange and amylose affinity chromatography
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recombinant N-terminal maltose-binding protein fusion enzyme from Escherichia coli strain BL21(DE3)RIPL by anion exchange and amylose affinity chromatography
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recombinant protein using His-tag
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recombinant Strep-tagged enzyme
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recombinant Strep-tagged wild-type and mutant enzyme by affinity chromatography to over 95% purity
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recombinant thioredoxin-His6-tagged enzyme by affinity chromatography
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recombinnat enzyme from Escherichia coli strain BL21(DE3) with cleavage of the Thx-His6 tag
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standard glutathione S-transferase fusion protein purification protocol, gel-filtration chromatography after removing the glutathione S-transferase tag
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the fusion protein is purified by immobilized metal (Ni2+) affinity chromatography. The liberated enzyme is separated from the His-8-maltose binding protein by immobilized metal affinity chromatography, further purified by gel filtration chromatography, and concentrated. EDTA is obmitted from the buffers used to purifiy enzyme for catalytic studies, and the gel filtration is not used.
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using acetone fractionation, column chromatography on DEAE-cellulose, Sephadex G-100, hydroxylapatite, DEAE-Sephadex A-25 and Sephadex G-75
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using acetone precipitation, first chromatography on DEAE-cellulose column, second chromatography on DEAE-cellulose column, chromatography on Sephadex G-100 column, hydroxyapatite column, DEAE-Sephadex A-25 column and Sephadex G-75 column
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using acid treatment, ammonium sulfate fractionation and column chromatography with DEAE-cellulose. The purified enzyme is composed of two distinct proteins, it appears that one of them is a catalytic protein named protein-B having tightly bound iron as a prosthetic group, while the other is either a modifier or activating protein named protein-A. Protein-B is found to exist in both an active and an inactive form
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using filtration, centrifugation, column chromatography on DEAE-cellulose, Sephadex G-50 and cysteine-Sepharose
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using heat treatment, ammonium sulfate fractionation and column chromatography on DEAE-cellulose, Sephadex G-200 and Sephadex G-100
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