Literature summary for 2.4.2.1 extracted from

Donaldson, T.M.; Ting, L.M.; Zhan, C.; Shi, W.; Zheng, R.; Almo, S.C.; Kim, K.
Structural determinants of the 5-methylthioinosine specificity of Plasmodium purine nucleoside phosphorylase (2014), PLoS ONE, 9, e84384.

Search for more literature for 2.4.2.1

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21(DE3) cells	Plasmodium falciparum

Crystallization (Commentary)

Crystallization (Comment)	Organism
mutant Y160F/V66I/V73I in complex with immucillin H, sitting drop vapor diffusion method, using 0.2 M magnesium chloride hexahydrate, 0.1 M HEPES at pH 7.5, and 30% (v/v) 2-propanol	Plasmodium falciparum

Protein Variants

Protein Variants	Comment	Organism
V66I	the mutant shows wild type activity	Plasmodium falciparum
V66I/V73I	the mutant shows increased activity compared to the wild type enzyme	Plasmodium falciparum
V66I/Y160F	the mutant shows increased activity compared to the wild type enzyme	Plasmodium falciparum
V73I	the mutant shows wild type activity	Plasmodium falciparum
V73I/Y160F	the mutant shows wild type activity	Plasmodium falciparum
Y160F	the mutant shows reduced activity compared to the wild type enzyme	Plasmodium falciparum
Y160F/V66I/V73I	the mutant has a 83fold decrease in turnover number for 5'-methylthioinosine with 2fold increase in Km value compared to the wild type enzyme	Plasmodium falciparum

Inhibitors

Inhibitors	Comment	Organism	Structure
5'-methylthio-immucillin H	-	Plasmodium falciparum
immucillin H	-	Plasmodium falciparum

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.0025	-	Inosine	mutant enzyme Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.0036	-	Inosine	mutant enzyme V73I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.004	-	Inosine	mutant enzyme V66I/Y73I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.0062	-	Inosine	mutant enzyme V66I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.0088	-	5'-methylthioinosine	wild type enzyme, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.011	-	Inosine	wild type enzyme, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.011	-	5'-methylthioinosine	mutant enzyme V66I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.011	-	5'-methylthioinosine	mutant enzyme V73I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.012	-	5'-methylthioinosine	mutant enzyme V66I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.013	-	Inosine	mutant enzyme V73I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.015	-	5'-methylthioinosine	mutant enzyme V66I/V73I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.016	-	Inosine	mutant enzyme V66I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.018	-	5'-methylthioinosine	mutant enzyme V66I/Y73I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.028	-	Inosine	mutant enzyme V66I/V73I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.044	-	5'-methylthioinosine	mutant enzyme Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.06	-	5'-methylthioinosine	mutant enzyme V73I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum

Organism

Organism	UniProt	Comment	Textmining
Plasmodium falciparum	Q8T9Z7	-	-

Purification (Commentary)

Purification (Comment)	Organism
Ni-NTA column chromatography	Plasmodium falciparum

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
5'-methylthioinosine + phosphate	-	Plasmodium falciparum	hypoxanthine + 5-methylthio-alpha-D-ribose 1-phosphate	-	r
guanosine + phosphate	-	Plasmodium falciparum	guanine + alpha-D-ribose 1-phosphate	-	?
inosine + phosphate	-	Plasmodium falciparum	hypoxanthine + alpha-D-ribose 1-phosphate	-	r

Subunits

Subunits	Comment	Organism
homohexamer	-	Plasmodium falciparum

Synonyms

Synonyms	Comment	Organism
PNP	-	Plasmodium falciparum
purine nucleoside phosphorylase	-	Plasmodium falciparum

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.01	-	5'-methylthioinosine	mutant enzyme V66I/Y73I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.05	-	5'-methylthioinosine	mutant enzyme V66I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.21	-	Inosine	mutant enzyme Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.28	-	5'-methylthioinosine	mutant enzyme V73I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.3	-	5'-methylthioinosine	mutant enzyme Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.36	-	Inosine	mutant enzyme V66I/Y73I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.59	-	Inosine	mutant enzyme V73I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.81	-	5'-methylthioinosine	mutant enzyme V73I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.83	-	5'-methylthioinosine	wild type enzyme, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.91	-	5'-methylthioinosine	mutant enzyme V66I/V73I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
1.2	-	5'-methylthioinosine	mutant enzyme V66I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
1.2	-	Inosine	mutant enzyme V66I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
1.7	-	Inosine	wild type enzyme, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
1.9	-	Inosine	mutant enzyme V73I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
2	-	Inosine	mutant enzyme V66I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
4.6	-	Inosine	mutant enzyme V66I/V73I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.0000018	-	5'-methylthio-immucillin H	pH 7.4, temperature not specified in the publication	Plasmodium falciparum
0.000015	-	immucillin H	pH 7.4, temperature not specified in the publication	Plasmodium falciparum

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.56	-	5'-methylthioinosine	mutant enzyme V66I/Y73I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
4.6	-	5'-methylthioinosine	mutant enzyme V73I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
4.7	-	5'-methylthioinosine	mutant enzyme V66I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
6.8	-	5'-methylthioinosine	mutant enzyme Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
62	-	5'-methylthioinosine	mutant enzyme V66I/V73I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
71	-	5'-methylthioinosine	mutant enzyme V73I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
83	-	Inosine	mutant enzyme Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
90	-	Inosine	mutant enzyme V66I/Y73I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
94	-	5'-methylthioinosine	mutant enzyme V66I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
95	-	5'-methylthioinosine	wild type enzyme, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
130	-	Inosine	mutant enzyme V66I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
140	-	Inosine	mutant enzyme V73I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
150	-	Inosine	wild type enzyme, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
160	-	Inosine	mutant enzyme V73I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
170	-	Inosine	mutant enzyme V66I/V73I, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum
190	-	Inosine	mutant enzyme V66I/Y160F, at pH 7.4, temperature not specified in the publication	Plasmodium falciparum

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Release 2023.1 (January 2023)