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Literature summary for 2.4.2.28 extracted from

  • Kang, X.; Zhao, Y.; Jiang, D.; Li, X.; Wang, X.; Wu, Y.; Chen, Z.; Zhang, X.C.
    Crystal structure and biochemical studies of Brucella melitensis 5-methylthioadenosine/S-adenosylhomocysteine nucleosidase (2014), Biochem. Biophys. Res. Commun., 446, 965-970.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Brucella melitensis

Crystallization (Commentary)

Crystallization (Comment) Organism
in complex with adenine, hanging drop vapor diffusion method, using 20% (w/v) PEG-1000, 0.1 M phosphate-citrate (pH 4.2) and 0.2 M Li2SO4 Brucella melitensis

Protein Variants

Protein Variants Comment Organism
A15M the mutant shows 14.5% catalytic efficiency compared to the wild type enzyme Brucella melitensis
D168A the mutation completely abolishes activity Brucella melitensis
D168N the mutation completely abolishes activity Brucella melitensis
E145A the mutation completely abolishes activity Brucella melitensis
E145Q the mutation completely abolishes activity Brucella melitensis
E18A the mutation completely abolishes activity Brucella melitensis
E18Q the mutation completely abolishes activity Brucella melitensis
M144A the mutant shows 0.4% catalytic efficiency compared to the wild type enzyme Brucella melitensis
R164A the mutant shows 15% catalytic efficiency compared to the wild type enzyme Brucella melitensis
R85A the mutant shows 6.3% catalytic efficiency compared to the wild type enzyme Brucella melitensis
R85H the mutant shows 7.1% catalytic efficiency compared to the wild type enzyme Brucella melitensis
S167A the mutant shows 3.3% catalytic efficiency compared to the wild type enzyme Brucella melitensis
V34A the mutant shows 12.5% catalytic efficiency compared to the wild type enzyme Brucella melitensis
V34I the mutant shows 13.9% catalytic efficiency compared to the wild type enzyme Brucella melitensis
W179A the mutant shows 3.3% catalytic efficiency compared to the wild type enzyme Brucella melitensis
W179F the mutant shows 2.0% catalytic efficiency compared to the wild type enzyme Brucella melitensis
Y134A the mutant shows 8.1% catalytic efficiency compared to the wild type enzyme Brucella melitensis
Y134F the mutant shows 0.6% catalytic efficiency compared to the wild type enzyme Brucella melitensis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0022
-
S-methyl-5'-thioadenosine wild type enzyme, at pH 7.0 and 25°C Brucella melitensis
0.0031
-
S-methyl-5'-thioadenosine mutant enzyme A15M, at pH 7.0 and 25°C Brucella melitensis
0.0044
-
S-methyl-5'-thioadenosine mutant enzyme S167A, at pH 7.0 and 25°C Brucella melitensis
0.0045
-
S-methyl-5'-thioadenosine mutant enzyme V34I, at pH 7.0 and 25°C Brucella melitensis
0.0048
-
S-methyl-5'-thioadenosine mutant enzyme R85H, at pH 7.0 and 25°C Brucella melitensis
0.0053
-
S-methyl-5'-thioadenosine mutant enzyme V34A, at pH 7.0 and 25°C Brucella melitensis
0.0054
-
S-methyl-5'-thioadenosine mutant enzyme R85A, at pH 7.0 and 25°C Brucella melitensis
0.0054
-
S-methyl-5'-thioadenosine mutant enzyme W179A, at pH 7.0 and 25°C Brucella melitensis
0.0074
-
S-methyl-5'-thioadenosine mutant enzyme W179F, at pH 7.0 and 25°C Brucella melitensis
0.028
-
S-methyl-5'-thioadenosine mutant enzyme Y134A, at pH 7.0 and 25°C Brucella melitensis
0.0301
-
S-methyl-5'-thioadenosine mutant enzyme Y134F, at pH 7.0 and 25°C Brucella melitensis
0.0351
-
S-methyl-5'-thioadenosine mutant enzyme M144A, at pH 7.0 and 25°C Brucella melitensis
8
-
S-methyl-5'-thioadenosine mutant enzyme R164A, at pH 7.0 and 25°C Brucella melitensis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
S-methyl-5'-thioadenosine + phosphate Brucella melitensis
-
adenine + S-methyl-5-thio-alpha-D-ribose 1-phosphate
-
ir

Organism

Organism UniProt Comment Textmining
Brucella melitensis Q8YBL1
-
-

Purification (Commentary)

Purification (Comment) Organism
nickel affinity column chromatography, and Superdex 200 gel filtration Brucella melitensis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
S-methyl-5'-thioadenosine + phosphate
-
Brucella melitensis adenine + S-methyl-5-thio-alpha-D-ribose 1-phosphate
-
ir

Synonyms

Synonyms Comment Organism
5'-methylthioadenosine/S-adenosylhomocysteine nucleosidase
-
Brucella melitensis
MTAN
-
Brucella melitensis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.51
-
S-methyl-5'-thioadenosine mutant enzyme M144A, at pH 7.0 and 25°C Brucella melitensis
0.58
-
S-methyl-5'-thioadenosine mutant enzyme S167A, at pH 7.0 and 25°C Brucella melitensis
0.58
-
S-methyl-5'-thioadenosine mutant enzyme W179F, at pH 7.0 and 25°C Brucella melitensis
0.69
-
S-methyl-5'-thioadenosine mutant enzyme W179A, at pH 7.0 and 25°C Brucella melitensis
0.71
-
S-methyl-5'-thioadenosine mutant enzyme Y134F, at pH 7.0 and 25°C Brucella melitensis
0.78
-
S-methyl-5'-thioadenosine mutant enzyme Y134A, at pH 7.0 and 25°C Brucella melitensis
1.3
-
S-methyl-5'-thioadenosine mutant enzyme R85H, at pH 7.0 and 25°C Brucella melitensis
1.4
-
S-methyl-5'-thioadenosine mutant enzyme R85A, at pH 7.0 and 25°C Brucella melitensis
1.8
-
S-methyl-5'-thioadenosine mutant enzyme A15M, at pH 7.0 and 25°C Brucella melitensis
2.5
-
S-methyl-5'-thioadenosine mutant enzyme V34I, at pH 7.0 and 25°C Brucella melitensis
2.7
-
S-methyl-5'-thioadenosine mutant enzyme V34A, at pH 7.0 and 25°C Brucella melitensis
8
-
S-methyl-5'-thioadenosine mutant enzyme R164A, at pH 7.0 and 25°C Brucella melitensis
8.8
-
S-methyl-5'-thioadenosine wild type enzyme, at pH 7.0 and 25°C Brucella melitensis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
15
-
S-methyl-5'-thioadenosine mutant enzyme M144A, at pH 7.0 and 25°C Brucella melitensis
24
-
S-methyl-5'-thioadenosine mutant enzyme Y134F, at pH 7.0 and 25°C Brucella melitensis
28
-
S-methyl-5'-thioadenosine mutant enzyme Y134A, at pH 7.0 and 25°C Brucella melitensis
79
-
S-methyl-5'-thioadenosine mutant enzyme W179F, at pH 7.0 and 25°C Brucella melitensis
130
-
S-methyl-5'-thioadenosine mutant enzyme S167A, at pH 7.0 and 25°C Brucella melitensis
130
-
S-methyl-5'-thioadenosine mutant enzyme W179A, at pH 7.0 and 25°C Brucella melitensis
250
-
S-methyl-5'-thioadenosine mutant enzyme R85A, at pH 7.0 and 25°C Brucella melitensis
280
-
S-methyl-5'-thioadenosine mutant enzyme R85H, at pH 7.0 and 25°C Brucella melitensis
510
-
S-methyl-5'-thioadenosine mutant enzyme V34A, at pH 7.0 and 25°C Brucella melitensis
560
-
S-methyl-5'-thioadenosine mutant enzyme V34I, at pH 7.0 and 25°C Brucella melitensis
580
-
S-methyl-5'-thioadenosine mutant enzyme A15M, at pH 7.0 and 25°C Brucella melitensis
600
-
S-methyl-5'-thioadenosine mutant enzyme R164A, at pH 7.0 and 25°C Brucella melitensis
4000
-
S-methyl-5'-thioadenosine wild type enzyme, at pH 7.0 and 25°C Brucella melitensis