Literature summary for 2.4.2.1 extracted from

Mikleusevic, G.; Stefanic, Z.; Narczyk, M.; Wielgus-Kutrowska, B.; Bzowska, A.; Luic, M.
Validation of the catalytic mechanism of Escherichia coli purine nucleoside phosphorylase by structural and kinetic studies (2011), Biochimie, 93, 1610-1622.

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli	Escherichia coli

Crystallization (Commentary)

Crystallization (Comment)	Organism
crystal structures of the wild type in complexes with phosphate and sulfate, respectively, and of the R24A mutant in complex with phosphate/sulfate, to about 2 A resolution. The structural data show that previously observed conformational change is a result of the phosphate binding and its interaction with residue Arg24	Escherichia coli

Protein Variants

Protein Variants	Comment	Organism
D204A	0.1% of wild-type activity towards inosine, adenosine and guanosine	Escherichia coli
D204A/R217A	about 0.2% of wild-type activity towards inosine, adenosine and guanosine	Escherichia coli
D204N	less than about 4% of wild-type activity towards inosine, adenosine and guanosine	Escherichia coli
R217A	about 0.5% of wild-type activity towards inosine and guanosine, 10.8% of activity towards adenosine	Escherichia coli
R24A	absence of a conformational change upon binding of phosphate, about 0.2 to 0.6% of wild-type activity towards inosine, adenosine and guanosine	Escherichia coli

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	P0ABP8	-	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.041	-	substrate inosine, mutant R217A, pH 7.0, 25°C	Escherichia coli
0.05	-	substrate adenosine, mutant D204A, pH 7.0, 25°C	Escherichia coli
0.06	-	substrate guanosine, mutant D204A, pH 7.0, 25°C	Escherichia coli
0.07	-	substrate adenosine, mutant D204N, pH 7.0, 25°C	Escherichia coli
0.07	-	substrate guanosine, mutant D204A/R217A, pH 7.0, 25°C	Escherichia coli
0.09	-	substrate guanosine, mutant R24A, pH 7.0, 25°C	Escherichia coli
0.09	-	substrate inosine, mutant D204A, pH 7.0, 25°C	Escherichia coli
0.13	-	substrate adenosine, mutant D204A/R217A, pH 7.0, 25°C	Escherichia coli
0.14	-	substrate inosine, mutant D204A/R217A, pH 7.0, 25°C	Escherichia coli
0.18	-	substrate inosine, mutant R24A, pH 7.0, 25°C	Escherichia coli
0.36	-	substrate adenosine, mutant R24A, pH 7.0, 25°C	Escherichia coli
0.36	-	substrate guanosine, mutant R217A, pH 7.0, 25°C	Escherichia coli
1.4	-	substrate inosine, mutant D204N, pH 7.0, 25°C	Escherichia coli
2.1	-	substrate guanosine, mutant D204N, pH 7.0, 25°C	Escherichia coli
6.3	-	substrate adenosine, mutant R217A, pH 7.0, 25°C	Escherichia coli
6.4	-	substrate 7-methylguanosine, mutant R24A, pH 7.0, 25°C	Escherichia coli
6.9	-	substrate 7-methylguanosine, mutant D204N, pH 7.0, 25°C	Escherichia coli
20	-	substrate 7-methylguanosine, mutant D204A, pH 7.0, 25°C	Escherichia coli
21.3	-	substrate 7-methylguanosine, mutant R217A, pH 7.0, 25°C	Escherichia coli
21.4	-	substrate 7-methylguanosine, wild-type, pH 7.0, 25°C	Escherichia coli
46.9	-	substrate 7-methylguanosine, mutant D204A/R217A, pH 7.0, 25°C	Escherichia coli
58.5	-	substrate adenosine, wild-type, pH 7.0, 25°C	Escherichia coli
60.4	-	substrate guanosine, wild-type, pH 7.0, 25°C	Escherichia coli
103.2	-	substrate inosine, wild-type, pH 7.0, 25°C	Escherichia coli

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
7-methylguanosine + phosphate	-	Escherichia coli	7-methylguanine + alpha-D-ribose 1-phosphate	-	?
adenosine + phosphate	-	Escherichia coli	adenine + alpha-D-ribose 1-phosphate	-	?
guanosine + phosphate	-	Escherichia coli	guanine + alpha-D-ribose 1-phosphate	-	?
inosine + phosphate	-	Escherichia coli	hypoxanthine + alpha-D-ribose 1-phosphate	-	?
additional information	catalysis involves protonation of the purine base at position N7 by residue Asp204, which is triggered by Arg217	Escherichia coli	?	-	?

Synonyms

Synonyms	Comment	Organism
DeoD	-	Escherichia coli