Literature summary for 3.5.4.2 extracted from

Kamat, S.S.; Bagaria, A.; Kumaran, D.; Holmes-Hampton, G.P.; Fan, H.; Sali, A.; Sauder, J.M.; Burley, S.K.; Lindahl, P.A.; Swaminathan, S.; Raushel, F.M.
Catalytic mechanism and three-dimensional structure of adenine deaminase (2011), Biochemistry, 50, 1917-1927.

Crystallization (Commentary)

Crystallization (Comment)	Organism
the three-dimensional structure of adenine deaminase from Agrobacterium tumefaciens (Atu4426) is determined by X-ray crystallography at 2.2 A resolution	Escherichia coli

Protein Variants

Protein Variants	Comment	Organism
D118N	mutant is able to bind 2 metals per active site, kcat (adenine): 173/sec	Escherichia coli
D284A	mutants is able to bind two equivalents of Mn2+ or Fe2+ in the active site but mutant is unable to catalyze the deaminase reaction	Escherichia coli
D285A	mutant is able to bind 2 metals per active site, kcat (adenine): 37/sec	Escherichia coli
D474N	mutant is able to bind 2 metals per active site, kcat (adenine): 171/sec	Escherichia coli
E121Q	mutant is able to bind 2 metals per active site, kcat (adenine): 57/sec	Escherichia coli
E185Q	mutant shows total loss of catalytic activity as well as the ability to bind divalent metals in the active site	Escherichia coli
E236Q	mutants is able to bind two equivalents of Mn2+ or Fe2+ in the active site but mutant is unable to catalyze the deaminase reaction	Escherichia coli
H120N	mutant is able to bind 2 metals per active site, kcat (adenine): 0.13/sec	Escherichia coli
H214N	mutant shows total loss of catalytic activity as well as the ability to bind divalent metals in the active site	Escherichia coli
H214Q	mutant shows total loss of catalytic activity as well as the ability to bind divalent metals in the active site	Escherichia coli
H235C	mutant shows total loss of catalytic activity as well as the ability to bind divalent metals in the active site	Escherichia coli
H235D	mutant shows total loss of catalytic activity as well as the ability to bind divalent metals in the active site	Escherichia coli
H235N	mutant shows total loss of catalytic activity as well as the ability to bind divalent metals in the active site	Escherichia coli
H235Q	mutant shows total loss of catalytic activity as well as the ability to bind divalent metals in the active site	Escherichia coli
H473N	mutant is able to bind 2 metals per active site, kcat (adenine): 178/sec	Escherichia coli
H90C	mutant is unable to bind either iron or manganese	Escherichia coli
H90D	mutant is unable to bind either iron or manganese	Escherichia coli
H90N	mutant is able to bind two equivalents of Mn2+ or Fe2+ per monomer and shows a kcat of about 5% of the wild-type enzyme	Escherichia coli
H90Q	mutant is unable to bind either iron or manganese	Escherichia coli
H92C	mutant shows total loss of catalytic activity as well as the ability to bind divalent metals in the active site	Escherichia coli
H92D	mutant shows total loss of catalytic activity as well as the ability to bind divalent metals in the active site	Escherichia coli
H92N	mutant shows total loss of catalytic activity as well as the ability to bind divalent metals in the active site	Escherichia coli
H92Q	mutant shows total loss of catalytic activity as well as the ability to bind divalent metals in the active site	Escherichia coli
S95A	mutant is able to bind 2 metals per active site, kcat (adenine): 78/sec	Escherichia coli

Inhibitors

Inhibitors	Comment	Organism	Structure
6-Chloropurine	-	Escherichia coli

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.23	-	adenine	pH 7.5, 30°C, wild-type Zn/Zn-ADE	Escherichia coli
0.3	-	adenine	pH 7.5, 30°C, wild-type Mn/Mn-ADE	Escherichia coli
0.31	-	adenine	mutant D474N, pH 7.5, 30°C	Escherichia coli
0.32	-	adenine	mutant D285A, pH 7.5, 30°C	Escherichia coli
0.32	-	adenine	mutant H214Q, pH 7.5, 30°C	Escherichia coli
0.32	-	adenine	mutant H473N, pH 7.5, 30°C	Escherichia coli
0.33	-	adenine	pH 7.5, 30°C, wild-type Fe/Fe-ADE	Escherichia coli
0.35	-	adenine	mutant D118N, pH 7.5, 30°C	Escherichia coli
0.35	-	adenine	mutant H90N, pH 7.5, 30°C	Escherichia coli
0.37	-	adenine	mutant H214N, pH 7.5, 30°C	Escherichia coli
0.38	-	adenine	mutant H92D, pH 7.5, 30°C	Escherichia coli
0.4	-	adenine	mutant H90Q, pH 7.5, 30°C	Escherichia coli
0.41	-	adenine	mutant E236Q, pH 7.5, 30°C	Escherichia coli
0.42	-	adenine	mutant H92N, pH 7.5, 30°C	Escherichia coli
0.44	-	adenine	mutant E121Q, pH 7.5, 30°C	Escherichia coli
0.44	-	adenine	mutant H90D, pH 7.5, 30°C	Escherichia coli
0.45	-	adenine	mutant H92C, pH 7.5, 30°C	Escherichia coli
0.47	-	adenine	mutant H120N, pH 7.5, 30°C	Escherichia coli
0.47	-	adenine	mutant S95A, pH 7.5, 30°C	Escherichia coli
0.48	-	adenine	mutant H235D, pH 7.5, 30°C	Escherichia coli
0.48	-	adenine	mutant H92Q, pH 7.5, 30°C	Escherichia coli
0.51	-	adenine	mutant H90C, pH 7.5, 30°C	Escherichia coli
0.8	-	adenine	mutant H235Q, pH 7.5, 30°C	Escherichia coli
0.82	-	adenine	mutant H235N, pH 7.5, 30°C	Escherichia coli
0.86	-	adenine	mutant D284A, pH 7.5, 30°C	Escherichia coli
1.2	-	adenine	mutant H235C, pH 7.5, 30°C	Escherichia coli
1.6	-	adenine	mutant E185Q, pH 7.5, 30°C	Escherichia coli

Metals/Ions

Metals/Ions	Comment	Organism
Fe2+	reducing [FeIII/FeIII]-ADE with dithionite restores the deaminase activity	Escherichia coli
Fe3+	[FeII/FeII]-ADE is oxidized to [FeIII/FeIII]-ADE with ferricyanide with inactivation of the deaminase activity	Escherichia coli
additional information	the apo-enzyme is prepared and reconstituted with Fe2+, Zn2+, or Mn2+. In each case, two enzyme-equivalents of metal are necessary for reconstitution of the deaminase activity	Escherichia coli

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	-	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
adenine + H2O	-	Escherichia coli	hypoxanthine + NH3	-	?

Synonyms

Synonyms	Comment	Organism
ADE	-	Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.022	-	adenine	mutant D284A, pH 7.5, 30°C	Escherichia coli
0.036	-	adenine	mutant H90D, pH 7.5, 30°C	Escherichia coli
0.05	-	adenine	mutant H92N, pH 7.5, 30°C	Escherichia coli
0.053	-	adenine	mutant H90C, pH 7.5, 30°C	Escherichia coli
0.06	-	adenine	mutant E185Q, pH 7.5, 30°C	Escherichia coli
0.072	-	adenine	mutant E236Q, pH 7.5, 30°C	Escherichia coli
0.08	-	adenine	mutant H92Q, pH 7.5, 30°C	Escherichia coli
0.13	-	adenine	mutant H120N, pH 7.5, 30°C	Escherichia coli
0.5	-	adenine	mutant H214N, pH 7.5, 30°C	Escherichia coli
0.5	-	adenine	mutant H235N, pH 7.5, 30°C	Escherichia coli
0.69	-	adenine	mutant H92C, pH 7.5, 30°C	Escherichia coli
0.8	-	adenine	mutant H90Q, pH 7.5, 30°C	Escherichia coli
0.8	-	adenine	mutant H92D, pH 7.5, 30°C	Escherichia coli
1.2	-	adenine	mutant H235D, pH 7.5, 30°C	Escherichia coli
1.3	-	adenine	mutant H214Q, pH 7.5, 30°C	Escherichia coli
1.3	-	adenine	mutant H235Q, pH 7.5, 30°C	Escherichia coli
1.5	-	adenine	mutant H235C, pH 7.5, 30°C	Escherichia coli
2	-	adenine	wild-type, pH 7.5, 30°C	Escherichia coli
8	-	adenine	mutant H90N, pH 7.5, 30°C	Escherichia coli
37	-	adenine	mutant D285A, pH 7.5, 30°C	Escherichia coli
57	-	adenine	mutant E121Q, pH 7.5, 30°C	Escherichia coli
78	-	adenine	mutant S95A, pH 7.5, 30°C	Escherichia coli
123	-	adenine	pH 7.5, 30°C, wild-type Zn/Zn-ADE	Escherichia coli
171	-	adenine	mutant D474N, pH 7.5, 30°C	Escherichia coli
173	-	adenine	mutant D118N, pH 7.5, 30°C	Escherichia coli
178	-	adenine	mutant H473N, pH 7.5, 30°C	Escherichia coli
185	-	adenine	pH 7.5, 30°C, wild-type Mn/Mn-ADE	Escherichia coli
196	-	adenine	pH 7.5, 30°C, wild-type Fe/Fe-ADE	Escherichia coli
200	-	adenine	wild-type, pH 7.5, 30°C, + Fe sequestered with a metal chelator, growth medium supplemented with Mn2+ prior to induction	Escherichia coli

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.00013	-	6-Chloropurine	pH 7.5, 30°C	Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.026	-	adenine	mutant D284A, pH 7.5, 30°C	Escherichia coli
0.038	-	adenine	mutant E185Q, pH 7.5, 30°C	Escherichia coli
0.082	-	adenine	mutant H90D, pH 7.5, 30°C	Escherichia coli
0.103	-	adenine	mutant H90C, pH 7.5, 30°C	Escherichia coli
0.119	-	adenine	mutant H92N, pH 7.5, 30°C	Escherichia coli
0.166	-	adenine	mutant H92Q, pH 7.5, 30°C	Escherichia coli
0.17	-	adenine	mutant E236Q, pH 7.5, 30°C	Escherichia coli
0.361	-	adenine	mutant H120N, pH 7.5, 30°C	Escherichia coli
0.609	-	adenine	mutant H235N, pH 7.5, 30°C	Escherichia coli
1.3	-	adenine	mutant H214N, pH 7.5, 30°C	Escherichia coli
1.3	-	adenine	mutant H235C, pH 7.5, 30°C	Escherichia coli
1.4	-	adenine	mutant H92C, pH 7.5, 30°C	Escherichia coli
1.6	-	adenine	mutant H235Q, pH 7.5, 30°C	Escherichia coli
2	-	adenine	mutant H90Q, pH 7.5, 30°C	Escherichia coli
2.1	-	adenine	mutant H92D, pH 7.5, 30°C	Escherichia coli
2.5	-	adenine	wild-type, pH 7.5, 30°C	Escherichia coli
2.5	-	adenine	mutant H235D, pH 7.5, 30°C	Escherichia coli
4	-	adenine	mutant H214Q, pH 7.5, 30°C	Escherichia coli
22	-	adenine	mutant H90N, pH 7.5, 30°C	Escherichia coli
130	-	adenine	mutant D285A, pH 7.5, 30°C	Escherichia coli
130	-	adenine	mutant E121Q, pH 7.5, 30°C	Escherichia coli
160	-	adenine	mutant S95A, pH 7.5, 30°C	Escherichia coli
500	-	adenine	wild-type, pH 7.5, 30°C, + Fe sequestered with a metal chelator, growth medium supplemented with Mn2+ prior to induction	Escherichia coli
540	-	adenine	pH 7.5, 30°C, wild-type Zn/Zn-ADE	Escherichia coli
550	-	adenine	mutant D474N, pH 7.5, 30°C	Escherichia coli
560	-	adenine	mutant D118N, pH 7.5, 30°C	Escherichia coli
560	-	adenine	mutant H473N, pH 7.5, 30°C	Escherichia coli
590	-	adenine	pH 7.5, 30°C, wild-type Fe/Fe-ADE	Escherichia coli
610	-	adenine	pH 7.5, 30°C, wild-type Mn/Mn-ADE	Escherichia coli