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Structural and functional analyses of Mycobacterium tuberculosis Rv3315c-encoded metal-dependent homotetrameric cytidine deaminase

Sanchez-Quitian, Z.A.; Schneider, C.Z.; Ducati, R.G.; de Azevedo, W.F.; Bloch, C.; Basso, L.A.; Santos, D.S.; J. Struct. Biol. 169, 413-423 (2010)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
3.5.4.5
expressed in Escherichia coli BL21(DE3) cells
Mycobacterium tuberculosis
Crystallization (Commentary)
EC Number
Crystallization
Organism
3.5.4.5
hanging drop vapor diffusion method
Mycobacterium tuberculosis
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.5.4.5
1.004
-
cytidine
in 50 mM Tris-HCl buffer pH 7.5 at 25°C
Mycobacterium tuberculosis
3.5.4.5
1.059
-
2'-deoxycytidine
in 50 mM Tris-HCl buffer pH 7.5 at 25°C
Mycobacterium tuberculosis
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
3.5.4.5
Zn2+
CDA is a zinc-dependent metalloenzyme and contains one mol of Zn2+ per mol of enzyme subunit
Mycobacterium tuberculosis
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.5.4.5
13938
-
4 * 13938, electrospray ionization mass spectrometry
Mycobacterium tuberculosis
3.5.4.5
14000
-
4 * 14000, SDS-PAGE
Mycobacterium tuberculosis
3.5.4.5
14072
-
4 * 14072, calculated from amino acid sequence
Mycobacterium tuberculosis
3.5.4.5
52990
-
gel filtration
Mycobacterium tuberculosis
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
3.5.4.5
Mycobacterium tuberculosis
P9WPH3
-
-
3.5.4.5
Mycobacterium tuberculosis H37Rv
P9WPH3
-
-
Purification (Commentary)
EC Number
Commentary
Organism
3.5.4.5
Q Sepharose column chromatography, Sephacryl S-200 gel filtration, and butyl-Sepharose column chromatography
Mycobacterium tuberculosis
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.5.4.5
1.68
-
crude extract, pH 7.5, at 25°C, using 2'-deoxycytidine as substrate
Mycobacterium tuberculosis
3.5.4.5
2.35
-
crude extract, pH 7.5, at 25°C, using cytidine as substrate
Mycobacterium tuberculosis
3.5.4.5
3.75
-
after 2.2fold purification, pH 7.5, at 25°C, using 2'-deoxycytidine as substrate
Mycobacterium tuberculosis
3.5.4.5
5.44
-
after 2.3fold purification, pH 7.5, at 25°C, using cytidine as substrate
Mycobacterium tuberculosis
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.5.4.5
2'-deoxycytidine + H2O
-
712889
Mycobacterium tuberculosis
2'-deoxyuridine + NH3
-
-
-
?
3.5.4.5
cytidine + H2O
-
712889
Mycobacterium tuberculosis
uridine + NH3
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
3.5.4.5
homotetramer
4 * 13938, electrospray ionization mass spectrometry; 4 * 14000, SDS-PAGE; 4 * 14072, calculated from amino acid sequence
Mycobacterium tuberculosis
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.5.4.5
3.5
-
2'-deoxycytidine
in 50 mM Tris-HCl buffer pH 7.5 at 25°C
Mycobacterium tuberculosis
3.5.4.5
4.8
-
cytidine
in 50 mM Tris-HCl buffer pH 7.5 at 25°C
Mycobacterium tuberculosis
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.4.5
expressed in Escherichia coli BL21(DE3) cells
Mycobacterium tuberculosis
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
3.5.4.5
hanging drop vapor diffusion method
Mycobacterium tuberculosis
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
3.5.4.5
1.004
-
cytidine
in 50 mM Tris-HCl buffer pH 7.5 at 25°C
Mycobacterium tuberculosis
3.5.4.5
1.059
-
2'-deoxycytidine
in 50 mM Tris-HCl buffer pH 7.5 at 25°C
Mycobacterium tuberculosis
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
3.5.4.5
Zn2+
CDA is a zinc-dependent metalloenzyme and contains one mol of Zn2+ per mol of enzyme subunit
Mycobacterium tuberculosis
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
3.5.4.5
13938
-
4 * 13938, electrospray ionization mass spectrometry
Mycobacterium tuberculosis
3.5.4.5
14000
-
4 * 14000, SDS-PAGE
Mycobacterium tuberculosis
3.5.4.5
14072
-
4 * 14072, calculated from amino acid sequence
Mycobacterium tuberculosis
3.5.4.5
52990
-
gel filtration
Mycobacterium tuberculosis
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
3.5.4.5
Q Sepharose column chromatography, Sephacryl S-200 gel filtration, and butyl-Sepharose column chromatography
Mycobacterium tuberculosis
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
3.5.4.5
1.68
-
crude extract, pH 7.5, at 25°C, using 2'-deoxycytidine as substrate
Mycobacterium tuberculosis
3.5.4.5
2.35
-
crude extract, pH 7.5, at 25°C, using cytidine as substrate
Mycobacterium tuberculosis
3.5.4.5
3.75
-
after 2.2fold purification, pH 7.5, at 25°C, using 2'-deoxycytidine as substrate
Mycobacterium tuberculosis
3.5.4.5
5.44
-
after 2.3fold purification, pH 7.5, at 25°C, using cytidine as substrate
Mycobacterium tuberculosis
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
3.5.4.5
2'-deoxycytidine + H2O
-
712889
Mycobacterium tuberculosis
2'-deoxyuridine + NH3
-
-
-
?
3.5.4.5
cytidine + H2O
-
712889
Mycobacterium tuberculosis
uridine + NH3
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
3.5.4.5
homotetramer
4 * 13938, electrospray ionization mass spectrometry; 4 * 14000, SDS-PAGE; 4 * 14072, calculated from amino acid sequence
Mycobacterium tuberculosis
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.5.4.5
3.5
-
2'-deoxycytidine
in 50 mM Tris-HCl buffer pH 7.5 at 25°C
Mycobacterium tuberculosis
3.5.4.5
4.8
-
cytidine
in 50 mM Tris-HCl buffer pH 7.5 at 25°C
Mycobacterium tuberculosis
General Information
EC Number
General Information
Commentary
Organism
3.5.4.5
metabolism
CDA is a pyrimidine salvage pathway enzyme that recycles cytidine and 20-deoxycytidine for uridine and 2’-deoxyuridine synthesis, respectively
Mycobacterium tuberculosis
General Information (protein specific)
EC Number
General Information
Commentary
Organism
3.5.4.5
metabolism
CDA is a pyrimidine salvage pathway enzyme that recycles cytidine and 20-deoxycytidine for uridine and 2’-deoxyuridine synthesis, respectively
Mycobacterium tuberculosis