Literature summary for 3.6.1.1 extracted from

Fabrichniy, I.P.; Lehtioe, L.; Tammenkoski, M.; Zyryanov, A.B.; Oksanen, E.; Baykov, A.A.; Lahti, R.; Goldman, A.
A trimetal site and substrate distortion in a family II inorganic pyrophosphatase (2007), J. Biol. Chem., 282, 1422-1431.

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant wild-type and mutant enzymes in complex with substrate analogue imidodiphosphate, PNP, and/or inhibitor fluoride, 35-40 mg/ml protein in 83mM TES/K+, pH 7.2, 17 mM KCl and 0.05 mM EGTA is mixed with 5 mM MgCl2 and 10 mM NaF, 1 mM PNP, sitting drop vapour diffusion method, 4°C, 3:2 ratio of protein to well solution, the latter containing 100 mM HEPES/K+, pH 7.5, 2.3-2.5 M ammonium sulfate, 3-4% PEG 400, 2-3 days, X-ray diffraction structure determination and anaylsis at 1.75-2.15 A resolution, the mutant H98Q crystals do not contain fluoride ions	Bacillus subtilis

Crystallization (Comment)

Organism

purified recombinant wild-type and mutant enzymes in complex with substrate analogue imidodiphosphate, PNP, and/or inhibitor fluoride, 35-40 mg/ml protein in 83mM TES/K+, pH 7.2, 17 mM KCl and 0.05 mM EGTA is mixed with 5 mM MgCl2 and 10 mM NaF, 1 mM PNP, sitting drop vapour diffusion method, 4°C, 3:2 ratio of protein to well solution, the latter containing 100 mM HEPES/K+, pH 7.5, 2.3-2.5 M ammonium sulfate, 3-4% PEG 400, 2-3 days, X-ray diffraction structure determination and anaylsis at 1.75-2.15 A resolution, the mutant H98Q crystals do not contain fluoride ions

Bacillus subtilis

Protein Variants

Protein Variants	Comment	Organism
H98Q	the mutant shows highly reduced activity compared to the wild-type enzyme, crystal structure determination and comparison of substrate/inhibitor binding to the wild-type enzyme	Bacillus subtilis

Inhibitors

Inhibitors	Comment	Organism	Structure
fluoride	inhibition of wild-type and mutant enzyme	Bacillus subtilis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.012	-	imidodiphosphoric acid	pH 7.2, 25°C, wild-type enzyme	Bacillus subtilis
0.06	-	diphosphate	pH 7.2, 25°C, wild-type enzyme	Bacillus subtilis

Metals/Ions

Metals/Ions	Comment	Organism
Fe3+	bound in in sites M1 and M2, the Fe3+:Mn2+ ratio is about 6:1 in site M1 and about 2:1 in site M2	Bacillus subtilis
Mn2+	bound in in sites M1 and M2, the Fe3+:Mn2+ ratio is about 6:1 in site M1 and about 2:1 in site M2	Bacillus subtilis
additional information	the enzyme contains a unique trinuclear metal center, detailed structure analysis, overview. Mn2+ and Fe3+ do not exchange for Mg2+ even in the presence of a large excess of Mg2+	Bacillus subtilis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
diphosphate + H2O	Bacillus subtilis	-	2 phosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Bacillus subtilis	P37487	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
diphosphate + H2O	-	Bacillus subtilis	2 phosphate	-	?
imidodiphosphate + H2O	active site and substrate binding structure determination and analysis, overview	Bacillus subtilis	phosphate + phosphoramidic acid	-	?

Synonyms

Synonyms	Comment	Organism
family II inorganic pyrophosphatase	-	Bacillus subtilis
family II PPase	-	Bacillus subtilis
PPase	-	Bacillus subtilis

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.014	-	imidodiphosphoric acid	pH 7.2, 25°C, wild-type enzyme	Bacillus subtilis

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.012	-	fluoride	pH 7.2, 25°C, wild-type enzyme, kinetics	Bacillus subtilis