Literature summary for 3.2.2.16 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.

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment)	Organism
in complex with its product 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 exhibits less than 16% catalytic efficiency compared to the wild type	Brucella melitensis
D168A	inactive	Brucella melitensis
D168N	inactive	Brucella melitensis
E145A	inactive	Brucella melitensis
E145Q	inactive	Brucella melitensis
E18A	inactive	Brucella melitensis
E18Q	inactive	Brucella melitensis
M144A	the mutant exhibits less than 1% catalytic efficiency compared to the wild type	Brucella melitensis
R164A	the variant displays catalytic efficiencies of less than 20% compared to the wild type	Brucella melitensis
R85A	the mutant exhibits less than 8% catalytic efficiency compared to the wild type	Brucella melitensis
R85H	the mutant exhibits less than 9% catalytic efficiency compared to the wild type	Brucella melitensis
S167A	the mutant exhibits less than 6% catalytic efficiency compared to the wild type	Brucella melitensis
V34A	the mutant exhibits less than 19% catalytic efficiency compared to the wild type	Brucella melitensis
V34I	the mutant exhibits less than 22% catalytic efficiency compared to the wild type	Brucella melitensis
W179A	the mutant exhibits less than 7% catalytic efficiency compared to the wild type	Brucella melitensis
W179F	the mutant exhibits less than 3% catalytic efficiency compared to the wild type	Brucella melitensis
Y134A	the mutant exhibits less than 1% catalytic efficiency compared to the wild type	Brucella melitensis
Y134F	the mutant exhibits less than 1% catalytic efficiency compared to the wild type	Brucella melitensis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.0022	-	S-methyl-5'-thioadenosine	wild type enzyme, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0031	-	S-methyl-5'-thioadenosine	mutant enzyme A15M, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.004	-	S-adenosyl-L-homocysteine	wild type enzyme, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0041	-	S-adenosyl-L-homocysteine	mutant enzyme A15M, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0044	-	S-methyl-5'-thioadenosine	mutant enzyme S167A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0045	-	S-methyl-5'-thioadenosine	mutant enzyme V34I, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0048	-	S-methyl-5'-thioadenosine	mutant enzyme R85H, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0053	-	S-methyl-5'-thioadenosine	mutant enzyme V34A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0054	-	S-methyl-5'-thioadenosine	mutant enzyme R85A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0054	-	S-methyl-5'-thioadenosine	mutant enzyme W179A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0056	-	S-adenosyl-L-homocysteine	mutant enzyme S167A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0058	-	S-adenosyl-L-homocysteine	mutant enzyme V34I, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0061	-	S-adenosyl-L-homocysteine	mutant enzyme R85H, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0062	-	S-adenosyl-L-homocysteine	mutant enzyme V34A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0068	-	S-adenosyl-L-homocysteine	mutant enzyme R85A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0074	-	S-methyl-5'-thioadenosine	mutant enzyme W179F, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0076	-	S-adenosyl-L-homocysteine	mutant enzyme W179A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.008	-	S-methyl-5'-thioadenosine	mutant enzyme R164A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0098	-	S-adenosyl-L-homocysteine	mutant enzyme W179F, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0133	-	S-adenosyl-L-homocysteine	mutant enzyme R164A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.028	-	S-methyl-5'-thioadenosine	mutant enzyme Y134A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0301	-	S-methyl-5'-thioadenosine	mutant enzyme Y134F, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0351	-	S-methyl-5'-thioadenosine	mutant enzyme M144A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0381	-	S-adenosyl-L-homocysteine	mutant enzyme Y134A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0452	-	S-adenosyl-L-homocysteine	mutant enzyme Y134F, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.0951	-	S-adenosyl-L-homocysteine	mutant enzyme M144A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
25000	-	2 * 25000, His-tagged enzyme, calculated from amino acid sequence	Brucella melitensis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
S-adenosyl-L-homocysteine + H2O	Brucella melitensis	-	S-D-ribosyl-L-homocysteine + adenine	-	ir
S-methyl-5'-thioadenosine + H2O	Brucella melitensis	-	S-methyl-5-thio-D-ribose + adenine	-	ir

Organism

Organism	UniProt	Comment	Textmining
Brucella melitensis	Q8YBL1	-	-

Purification (Commentary)

Purification (Comment)	Organism
ammonium sulfate precipitation, 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-adenosyl-L-homocysteine + H2O	-	Brucella melitensis	S-D-ribosyl-L-homocysteine + adenine	-	ir
S-methyl-5'-thioadenosine + H2O	-	Brucella melitensis	S-methyl-5-thio-D-ribose + adenine	-	ir

Subunits

Subunits	Comment	Organism
homodimer	2 * 25000, His-tagged enzyme, calculated from amino acid sequence	Brucella melitensis

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
0.15	-	S-adenosyl-L-homocysteine	mutant enzyme M144A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.35	-	S-adenosyl-L-homocysteine	mutant enzyme Y134F, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.37	-	S-adenosyl-L-homocysteine	mutant enzyme W179F, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.39	-	S-adenosyl-L-homocysteine	mutant enzyme Y134A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.51	-	S-methyl-5'-thioadenosine	mutant enzyme M144A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.55	-	S-adenosyl-L-homocysteine	mutant enzyme S167A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.58	-	S-methyl-5'-thioadenosine	mutant enzyme S167A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.58	-	S-methyl-5'-thioadenosine	mutant enzyme W179F, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.69	-	S-methyl-5'-thioadenosine	mutant enzyme W179A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.71	-	S-methyl-5'-thioadenosine	mutant enzyme Y134F, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.78	-	S-methyl-5'-thioadenosine	mutant enzyme Y134A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.86	-	S-adenosyl-L-homocysteine	mutant enzyme W179A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.92	-	S-adenosyl-L-homocysteine	mutant enzyme R85H, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
0.93	-	S-adenosyl-L-homocysteine	mutant enzyme R85A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
1.1	-	S-adenosyl-L-homocysteine	mutant enzyme A15M, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
1.3	-	S-methyl-5'-thioadenosine	mutant enzyme R85H, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
1.4	-	S-methyl-5'-thioadenosine	mutant enzyme R85A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
1.8	-	S-methyl-5'-thioadenosine	mutant enzyme A15M, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
2.1	-	S-adenosyl-L-homocysteine	mutant enzyme V34A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
2.3	-	S-adenosyl-L-homocysteine	mutant enzyme V34I, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
2.5	-	S-methyl-5'-thioadenosine	mutant enzyme V34I, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
2.7	-	S-methyl-5'-thioadenosine	mutant enzyme V34A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
4.7	-	S-adenosyl-L-homocysteine	mutant enzyme R164A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
7.1	-	S-adenosyl-L-homocysteine	wild type enzyme, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
8	-	S-methyl-5'-thioadenosine	mutant enzyme R164A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
8.8	-	S-methyl-5'-thioadenosine	wild type enzyme, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
1.6	-	S-adenosyl-L-homocysteine	mutant enzyme M144A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
7.8	-	S-adenosyl-L-homocysteine	mutant enzyme Y134F, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
10	-	S-adenosyl-L-homocysteine	mutant enzyme Y134A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
15	-	S-methyl-5'-thioadenosine	mutant enzyme M144A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
24	-	S-methyl-5'-thioadenosine	mutant enzyme Y134F, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
28	-	S-methyl-5'-thioadenosine	mutant enzyme Y134A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
38	-	S-adenosyl-L-homocysteine	mutant enzyme W179F, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
79	-	S-methyl-5'-thioadenosine	mutant enzyme W179F, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
100	-	S-adenosyl-L-homocysteine	mutant enzyme S167A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
110	-	S-adenosyl-L-homocysteine	mutant enzyme W179A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
130	-	S-methyl-5'-thioadenosine	mutant enzyme S167A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
130	-	S-methyl-5'-thioadenosine	mutant enzyme W179A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
140	-	S-adenosyl-L-homocysteine	mutant enzyme R85A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
150	-	S-adenosyl-L-homocysteine	mutant enzyme R85H, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
250	-	S-methyl-5'-thioadenosine	mutant enzyme R85A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
270	-	S-adenosyl-L-homocysteine	mutant enzyme A15M, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
280	-	S-methyl-5'-thioadenosine	mutant enzyme R85H, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
330	-	S-adenosyl-L-homocysteine	mutant enzyme V34A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
350	-	S-adenosyl-L-homocysteine	mutant enzyme R164A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
390	-	S-adenosyl-L-homocysteine	mutant enzyme V34I, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
510	-	S-methyl-5'-thioadenosine	mutant enzyme V34A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
560	-	S-methyl-5'-thioadenosine	mutant enzyme V34I, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
580	-	S-methyl-5'-thioadenosine	mutant enzyme A15M, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
600	-	S-methyl-5'-thioadenosine	mutant enzyme R164A, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
1800	-	S-adenosyl-L-homocysteine	wild type enzyme, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis
4000	-	S-methyl-5'-thioadenosine	wild type enzyme, in 100 mM HEPES (pH 7.5), at 25°C	Brucella melitensis