Literature summary for 3.1.3.5 extracted from

Allegrini, S.; Scaloni, A.; Careddu, M.G.; Cuccu, G.; D'Ambrosio, C.; Pesi, R.; Camici, M.; Ferrara, L.; Tozzi, M.G.
Mechanistic studies on bovine cytosolic 5'-nucleotidase II, an enzyme belonging to the HAD superfamily (2004), Eur. J. Biochem., 271, 4881-4891.

Protein Variants

Protein Variants	Comment	Organism
D351E	strong decrease in nucleotidase activity, strong reduction in affinity towards Mg2+	Bos taurus
D356E	strong decrease in nucleotidase activity, strong reduction in affinity towards Mg2+	Bos taurus
D356N	strong increase in Km-value for inosine	Bos taurus
K292M	strong decrease in nucleotidase activity	Bos taurus
K292R	strong decrease in nucleotidase activity	Bos taurus
M53I	lowest nucleotidase vs. phosphotransferase activity ratio of all mutants tested	Bos taurus
M53N	strong decrease in catalytic efficiency	Bos taurus
S251A	kinetic behaviour almost similar to wild-type	Bos taurus
S251T	kinetic behaviour almost similar to wild-type	Bos taurus
T249S	kinetic behaviour almost similar to wild-type	Bos taurus
T249V	strongly reduced enzyme activity	Bos taurus
T56R	mutant devoid of phosphotransferase and nucleotidase activities	Bos taurus

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.1	-	5'-IMP	wild-type	Bos taurus
0.1	-	5'-IMP	mutant M53N	Bos taurus
0.1	-	5'-IMP	mutant S251A	Bos taurus
0.2	-	5'-IMP	mutant D356E	Bos taurus
0.2	-	5'-IMP	mutant D356N	Bos taurus
0.2	-	5'-IMP	mutant T249S	Bos taurus
0.25	-	Inosine	mutant T249v	Bos taurus
0.3	-	5'-IMP	mutant S251T	Bos taurus
0.4	-	5'-IMP	mutant T249v	Bos taurus
0.5	-	Inosine	mutant K292M	Bos taurus
0.8	-	Inosine	mutant M53I	Bos taurus
0.9	-	Inosine	mutant D351E	Bos taurus
0.9	-	Inosine	mutant T249S	Bos taurus
1	-	Inosine	mutant D356E	Bos taurus
1	-	5'-IMP	mutant M53I	Bos taurus
1	-	Inosine	mutant S251A	Bos taurus
1.1	-	Inosine	wild-type	Bos taurus
1.1	-	Inosine	mutant M53N	Bos taurus
1.5	-	Inosine	mutant S251T	Bos taurus
10.2	-	Inosine	mutant D356N	Bos taurus

Localization	Comment	Organism	GeneOntology No.	Textmining
cytosol	-	Bos taurus	5829	-

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Bos taurus

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
60000	-	x * 60000, SDS-PAGE	Bos taurus

Organism	UniProt	Comment	Textmining
Bos taurus	-	isoform 5'-nucleotidase II	-

Reaction	Comment	Organism	Reaction ID
a 5'-ribonucleotide + H2O = a ribonucleoside + phosphate	conserved motif I is involved in formation and stabilization of the covalent enzyme-phosphate intermediate. T249 and K292 in motif II contribute to stabilize the enzyme-phospho adduct. D351 and D356 in motif III coordinate the magnesium ion required for catalysis	Bos taurus	a

Renatured (Comment)	Organism
inhibition by CuCl2, 5,5-dithiobis-(2-nitro-benzoic acid) is reverted by presence of reducing agent	Bos taurus

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
5'-IMP + H2O	-	Bos taurus	inosine + phosphate	-	r
inosine + phosphate	-	Bos taurus	5'-IMP + H2O	-	r

Subunits	Comment	Organism
?	x * 60000, SDS-PAGE	Bos taurus

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.1	-	Inosine	mutant D351E	Bos taurus
0.1	-	Inosine	mutant K292M	Bos taurus
0.3	-	Inosine	mutant T249v	Bos taurus
0.6	-	Inosine	mutant M53N	Bos taurus
1.4	-	Inosine	mutant D356N	Bos taurus
2.2	-	Inosine	mutant D356E	Bos taurus
3.4	-	Inosine	mutant S251T	Bos taurus
3.7	-	Inosine	mutant M53I	Bos taurus
12.3	-	Inosine	mutant S251A	Bos taurus
19.8	-	Inosine	mutant T249S	Bos taurus
40	-	Inosine	wild-type	Bos taurus