Literature summary for 1.13.11.20 extracted from

Simmons, C.R.; Hirschberger, L.L.; Machi, M.S.; Stipanuk, M.H.
Expression, purification, and kinetic characterization of recombinant rat cysteine dioxygenase, a non-heme metalloenzyme necessary for regulation of cellular cysteine levels (2006), Protein Expr. Purif., 47, 74-81.

Search for more literature for 1.13.11.20

Activating Compound

Activating Compound	Comment	Organism	Structure
additional information	with pure cysteine dioxygenase no effect of addition of NAD+ to the cysteine diosygenase is found.	Rattus norvegicus

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli strain BL21(DE3)	Rattus norvegicus

Protein Variants

Protein Variants	Comment	Organism
additional information	recombinant cysteine dioxygenase protein with a thioredoxin sequence and 6 x His tagged, after purification the 6x His tag and the thioredoxin sequence are removed using factor Xa.	Rattus norvegicus

Inhibitors

Inhibitors	Comment	Organism	Structure
L-cysteine	concentrations of cysteine of 2 mM and above are inhibitory in assays of purified cysteine dioxygenase	Rattus norvegicus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.45	-	L-cysteine	for purified protein	Rattus norvegicus

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Fe2+	additon of 0.2 or 0.3 mM Fe2+ yields near-optimal activity. Activity of fully purified enzyme in the absence of added Fe2+ is below 1% of that measured in the presence of 0.3 mM ferrous sulfate.	Rattus norvegicus

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
23030	-	MALDI-MS	Rattus norvegicus
23030	-	theoretical value, calculated from the primary sequence	Rattus norvegicus

Organism

Organism	UniProt	Comment	Textmining
Rattus norvegicus	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
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.	Rattus norvegicus

Purification (Comment)

Organism

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.

Rattus norvegicus

Source Tissue

Source Tissue	Comment	Organism	Textmining
liver	cDNA	Rattus norvegicus	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
additional information	-	With Lineweaver-Burk plot Vmax is estimated to be 1.870 micromol cysteinesulfinate/min/mg	Rattus norvegicus
1.357	-	-	Rattus norvegicus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
L-cysteine + O2	-	Rattus norvegicus	3-sulfino-L-alanine	-	?

Synonyms

Synonyms	Comment	Organism
CDO	-	Rattus norvegicus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.012	-	L-cysteine	assay standart conditions are: the enzyme is incubated at 37°C in the presence of 62.5 mM Mes buffer (pH 6.1), 0.3 mM ferrous sulfate, 0.0125 mM bathocuproine disulfonate and 1.2 mM cysteine	Rattus norvegicus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
5.8	6.2	highest activity	Rattus norvegicus