Literature summary for 3.2.2.9 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.

Search for more literature for 3.2.2.9

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 phosphatecitrate (pH 4.2) and 0.2 M Li2SO4	Brucella melitensis

Protein Variants

Protein Variants	Comment	Organism
D168A	completely inactive	Brucella melitensis
D168N	completely inactive	Brucella melitensis
E145Q	completely inactive	Brucella melitensis
E18A	completely inactive	Brucella melitensis
E18Q	completely inactive	Brucella melitensis
M144A	the mutant shows 0.1% catalytic efficiency compared to the wild type enzyme	Brucella melitensis
R164A	the mutant shows 19.4% catalytic efficiency compared to the wild type enzyme	Brucella melitensis
R85A	the mutant shows 7.6% catalytic efficiency compared to the wild type enzyme	Brucella melitensis
R85H	the mutant shows 8.1% catalytic efficiency compared to the wild type enzyme	Brucella melitensis
S167A	the mutant shows 5.6% catalytic efficiency compared to the wild type enzyme	Brucella melitensis
Y134A	the mutant shows 0.6% catalytic efficiency compared to the wild type enzyme	Brucella melitensis
Y134F	the mutant shows 0.4% catalytic efficiency compared to the wild type enzyme	Brucella melitensis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.004	-	S-adenosyl-L-homocysteine	wild type enzyme, at pH 7.0 and 37°C	Brucella melitensis
0.0056	-	S-adenosyl-L-homocysteine	mutant enzyme S167A, at pH 7.0 and 37°C	Brucella melitensis
0.0061	-	S-adenosyl-L-homocysteine	mutant enzyme R85H, at pH 7.0 and 37°C	Brucella melitensis
0.0068	-	S-adenosyl-L-homocysteine	mutant enzyme R85A, at pH 7.0 and 37°C	Brucella melitensis
0.0133	-	S-adenosyl-L-homocysteine	mutant enzyme R164A, at pH 7.0 and 37°C	Brucella melitensis
0.0381	-	S-adenosyl-L-homocysteine	mutant enzyme Y134A, at pH 7.0 and 37°C	Brucella melitensis
0.0452	-	S-adenosyl-L-homocysteine	mutant enzyme Y134F, at pH 7.0 and 37°C	Brucella melitensis
0.0951	-	S-adenosyl-L-homocysteine	mutant enzyme M144A, at pH 7.0 and 37°C	Brucella melitensis

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
25000	-	x * 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-(5-deoxy-D-ribos-5-yl)-L-homocysteine + adenine	-	?

Organism

Organism	UniProt	Comment	Textmining
Brucella melitensis	-	-	-

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-adenosyl-L-homocysteine + H2O	-	Brucella melitensis	S-(5-deoxy-D-ribos-5-yl)-L-homocysteine + adenine	-	?

Subunits

Subunits	Comment	Organism
?	x * 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	Structure
0.15	-	S-adenosyl-L-homocysteine	mutant enzyme M144A, at pH 7.0 and 37°C	Brucella melitensis
0.35	-	S-adenosyl-L-homocysteine	mutant enzyme Y134F, at pH 7.0 and 37°C	Brucella melitensis
0.39	-	S-adenosyl-L-homocysteine	mutant enzyme Y134A, at pH 7.0 and 37°C	Brucella melitensis
0.55	-	S-adenosyl-L-homocysteine	mutant enzyme S167A, at pH 7.0 and 37°C	Brucella melitensis
0.92	-	S-adenosyl-L-homocysteine	mutant enzyme R85H, at pH 7.0 and 37°C	Brucella melitensis
0.93	-	S-adenosyl-L-homocysteine	mutant enzyme R85A, at pH 7.0 and 37°C	Brucella melitensis
4.7	-	S-adenosyl-L-homocysteine	mutant enzyme R164A, at pH 7.0 and 37°C	Brucella melitensis
7.1	-	S-adenosyl-L-homocysteine	wild type enzyme, at pH 7.0 and 37°C	Brucella melitensis

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
1.6	-	S-adenosyl-L-homocysteine	mutant enzyme M144A, at pH 7.0 and 37°C	Brucella melitensis
7.8	-	S-adenosyl-L-homocysteine	mutant enzyme Y134F, at pH 7.0 and 37°C	Brucella melitensis
10	-	S-adenosyl-L-homocysteine	mutant enzyme Y134A, at pH 7.0 and 37°C	Brucella melitensis
100	-	S-adenosyl-L-homocysteine	mutant enzyme S167A, at pH 7.0 and 37°C	Brucella melitensis
140	-	S-adenosyl-L-homocysteine	mutant enzyme R85A, at pH 7.0 and 37°C	Brucella melitensis
150	-	S-adenosyl-L-homocysteine	mutant enzyme R85H, at pH 7.0 and 37°C	Brucella melitensis
350	-	S-adenosyl-L-homocysteine	mutant enzyme R164A, at pH 7.0 and 37°C	Brucella melitensis
1800	-	S-adenosyl-L-homocysteine	wild type enzyme, at pH 7.0 and 37°C	Brucella melitensis

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Release 2023.1 (January 2023)