Literature summary for 3.1.3.1 extracted from

Hehir, M.J.; Murphy, J.E.; Kantrowitz, E.R.
Characterization of heterodimeric alkaline phosphatases from Escherichia coli: An Investigation of Intragenic Complementation (2000), J. Mol. Biol., 304, 645-656.

Search for more literature for 3.1.3.1

Activating Compound

Activating Compound	Comment	Organism	Structure
Mg2+	-	Escherichia coli
Zn2+	stimulation of mutant enzymes, in presence or absence of a phosphoryl group acceptor in the reaction, except for mutnat H412Y, whose activity is reduced in presence of phosphoryl group acceptor and 0.1 mM Zn2+	Escherichia coli

Cloned(Commentary)

Cloned (Comment)	Organism
expression of wild-type and mutants in several Escherichia coli strains, intragenic complementation	Escherichia coli

Protein Variants

Protein Variants	Comment	Organism
D369N	site-directed mutagenesis, highly reduced activity	Escherichia coli
D434E	site-directed mutagenesis, reduced activity, increased kcat and Km compared to the wild-type enzyme	Escherichia coli
E322K	site-directed mutagenesis, highly reduced activity	Escherichia coli
E341K	site-directed mutagenesis, highly reduced activity	Escherichia coli
H412Y	site-directed mutagenesis, highly reduced activity	Escherichia coli
S105L	site-directed mutagenesis, reduced activity	Escherichia coli
T155M	site-directed mutagenesis, highly reduced activity	Escherichia coli

Inhibitors

Inhibitors	Comment	Organism	Structure
Zn2+	mutant H412Y in presence of a phosphoryl group acceptor and Zn2+ at 0.1 mM	Escherichia coli

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	kinetic characterization of the homodimeric complementation mutants, and heterodimeric hybrid formation	Escherichia coli
0.0035	-	4-nitrophenyl phosphate	mutant S105L, pH 8.0, 25°C, in absence of a phosphoryl group acceptor	Escherichia coli
0.005	-	4-nitrophenyl phosphate	mutant T155M, pH 8.0, 25°C, in absence of a phosphoryl group acceptor	Escherichia coli
0.0073	-	4-nitrophenyl phosphate	mutant H412Y, pH 8.0, 25°C, in absence of a phosphoryl group acceptor	Escherichia coli
0.0094	-	4-nitrophenyl phosphate	wild-type enzyme, pH 8.0, 25°C, in absence of a phosphoryl group acceptor	Escherichia coli
0.0095	-	4-nitrophenyl phosphate	mutant E341K, pH 8.0, 25°C, in absence of a phosphoryl group acceptor	Escherichia coli
0.0097	-	4-nitrophenyl phosphate	mutant S105L, pH 8.0, 25°C, in presence of a phosphoryl group acceptor	Escherichia coli
0.011	-	4-nitrophenyl phosphate	mutant E322K, pH 8.0, 25°C, in presence of a phosphoryl group acceptor	Escherichia coli
0.0176	-	4-nitrophenyl phosphate	mutant D369N, pH 8.0, 25°C, in absence of a phosphoryl group acceptor	Escherichia coli
0.0211	-	4-nitrophenyl phosphate	wild-type enzyme, pH 8.0, 25°C, in presence of a phosphoryl group acceptor	Escherichia coli
0.0221	-	4-nitrophenyl phosphate	mutant E322K, pH 8.0, 25°C, in absence of a phosphoryl group acceptor	Escherichia coli
0.0374	-	4-nitrophenyl phosphate	mutant E341K, pH 8.0, 25°C, in presence of a phosphoryl group acceptor	Escherichia coli
0.057	-	4-nitrophenyl phosphate	mutant T155M, pH 8.0, 25°C, in presence of a phosphoryl group acceptor	Escherichia coli
0.0597	-	4-nitrophenyl phosphate	mutant D369N, pH 8.0, 25°C, in presence of a phosphoryl group acceptor	Escherichia coli
0.0691	-	4-nitrophenyl phosphate	mutant D434E, pH 8.0, 25°C, in absence of a phosphoryl group acceptor	Escherichia coli
0.198	-	4-nitrophenyl phosphate	mutant H412Y, pH 8.0, 25°C, in presence of a phosphoryl group acceptor	Escherichia coli

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	-	gene phoA	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type and mutants, isolation of heterodimeric forms	Escherichia coli

Reaction

Reaction	Comment	Organism	Reaction ID
a phosphate monoester + H2O = an alcohol + phosphate	Wide specificity. Also catalyses transphosphorylations. Some enzymes hydrolyse diphosphate	Escherichia coli	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
4-nitrophenyl phosphate + H2O	-	Escherichia coli	4-nitrophenol + phosphate	-	?

Subunits

Subunits	Comment	Organism
More	heterodimer formation in vitro, overview, structural assembly of the enzyme into dimer induces cooperative interactions between the monomers necessary for the formation of the functional form of the holoenzyme	Escherichia coli

Synonyms

Synonyms	Comment	Organism
alkaline phosphatase	-	Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
additional information	-	additional information	-	Escherichia coli
0.016	-	4-nitrophenyl phosphate	mutant E322K, pH 8.0, 25°C, in absence of a phosphoryl group acceptor	Escherichia coli
0.027	-	4-nitrophenyl phosphate	mutant E322K, pH 8.0, 25°C, in presence of a phosphoryl group acceptor	Escherichia coli
0.042	-	4-nitrophenyl phosphate	mutant E341K, pH 8.0, 25°C, in absence of a phosphoryl group acceptor	Escherichia coli
0.052	-	4-nitrophenyl phosphate	mutant T155M, pH 8.0, 25°C, in absence of a phosphoryl group acceptor	Escherichia coli
0.059	-	4-nitrophenyl phosphate	mutant H412Y, pH 8.0, 25°C, in presence of a phosphoryl group acceptor	Escherichia coli
0.067	-	4-nitrophenyl phosphate	mutant H412Y, pH 8.0, 25°C, in absence of a phosphoryl group acceptor	Escherichia coli
0.1	-	4-nitrophenyl phosphate	mutant E341K, pH 8.0, 25°C, in presence of a phosphoryl group acceptor	Escherichia coli
0.32	-	4-nitrophenyl phosphate	mutant T155M, pH 8.0, 25°C, in presence of a phosphoryl group acceptor	Escherichia coli
0.39	-	4-nitrophenyl phosphate	mutant D369N, pH 8.0, 25°C, in absence of a phosphoryl group acceptor	Escherichia coli
2.4	-	4-nitrophenyl phosphate	mutant D369N, pH 8.0, 25°C, in presence of a phosphoryl group acceptor	Escherichia coli
3.8	-	4-nitrophenyl phosphate	mutant S105L, pH 8.0, 25°C, in absence of a phosphoryl group acceptor	Escherichia coli
5.8	-	4-nitrophenyl phosphate	mutant S105L, pH 8.0, 25°C, in presence of a phosphoryl group acceptor	Escherichia coli
44.4	-	4-nitrophenyl phosphate	wild-type enzyme, pH 8.0, 25°C, in absence of a phosphoryl group acceptor	Escherichia coli
65.6	-	4-nitrophenyl phosphate	mutant D434E, pH 8.0, 25°C, in absence of a phosphoryl group acceptor	Escherichia coli
80.5	-	4-nitrophenyl phosphate	wild-type enzyme, pH 8.0, 25°C, in presence of a phosphoryl group acceptor	Escherichia coli

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Release 2023.1 (January 2023)