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Literature summary 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.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

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

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.4.2.1 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

EC Number Protein Variants Comment Organism
2.4.2.1 V66I the mutant shows wild type activity Plasmodium falciparum
2.4.2.1 V66I/V73I the mutant shows increased activity compared to the wild type enzyme Plasmodium falciparum
2.4.2.1 V66I/Y160F the mutant shows increased activity compared to the wild type enzyme Plasmodium falciparum
2.4.2.1 V73I the mutant shows wild type activity Plasmodium falciparum
2.4.2.1 V73I/Y160F the mutant shows wild type activity Plasmodium falciparum
2.4.2.1 Y160F the mutant shows reduced activity compared to the wild type enzyme Plasmodium falciparum
2.4.2.1 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

EC Number Inhibitors Comment Organism Structure
2.4.2.1 5'-methylthio-immucillin H
-
 Plasmodium falciparum
2.4.2.1 immucillin H
-
 Plasmodium falciparum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.2.1 0.0025
-
 Inosine mutant enzyme Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.0036
-
 Inosine mutant enzyme V73I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.004
-
 Inosine mutant enzyme V66I/Y73I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.0062
-
 Inosine mutant enzyme V66I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.0088
-
 5'-methylthioinosine wild type enzyme, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.011
-
 Inosine wild type enzyme, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.011
-
 5'-methylthioinosine mutant enzyme V66I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.011
-
 5'-methylthioinosine mutant enzyme V73I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.012
-
 5'-methylthioinosine mutant enzyme V66I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.013
-
 Inosine mutant enzyme V73I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.015
-
 5'-methylthioinosine mutant enzyme V66I/V73I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.016
-
 Inosine mutant enzyme V66I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.018
-
 5'-methylthioinosine mutant enzyme V66I/Y73I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.028
-
 Inosine mutant enzyme V66I/V73I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.044
-
 5'-methylthioinosine mutant enzyme Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.06
-
 5'-methylthioinosine mutant enzyme V73I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum

Organism

EC Number Organism UniProt Comment Textmining
2.4.2.1 Plasmodium falciparum Q8T9Z7
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.2.1 5'-methylthioinosine + phosphate
-
 Plasmodium falciparum hypoxanthine + 5-methylthio-alpha-D-ribose 1-phosphate
-
 r
2.4.2.1 guanosine + phosphate
-
 Plasmodium falciparum guanine + alpha-D-ribose 1-phosphate
-
 ?
2.4.2.1 inosine + phosphate
-
 Plasmodium falciparum hypoxanthine + alpha-D-ribose 1-phosphate
-
 r

Subunits

EC Number Subunits Comment Organism
2.4.2.1 homohexamer
-
 Plasmodium falciparum

Synonyms

EC Number Synonyms Comment Organism
2.4.2.1 PNP
-
 Plasmodium falciparum
2.4.2.1 purine nucleoside phosphorylase
-
 Plasmodium falciparum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.2.1 0.01
-
 5'-methylthioinosine mutant enzyme V66I/Y73I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.05
-
 5'-methylthioinosine mutant enzyme V66I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.21
-
 Inosine mutant enzyme Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.28
-
 5'-methylthioinosine mutant enzyme V73I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.3
-
 5'-methylthioinosine mutant enzyme Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.36
-
 Inosine mutant enzyme V66I/Y73I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.59
-
 Inosine mutant enzyme V73I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.81
-
 5'-methylthioinosine mutant enzyme V73I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.83
-
 5'-methylthioinosine wild type enzyme, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.91
-
 5'-methylthioinosine mutant enzyme V66I/V73I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 1.2
-
 5'-methylthioinosine mutant enzyme V66I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 1.2
-
 Inosine mutant enzyme V66I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 1.7
-
 Inosine wild type enzyme, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 1.9
-
 Inosine mutant enzyme V73I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 2
-
 Inosine mutant enzyme V66I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 4.6
-
 Inosine mutant enzyme V66I/V73I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.4.2.1 0.0000018
-
 5'-methylthio-immucillin H pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 0.000015
-
 immucillin H pH 7.4, temperature not specified in the publication Plasmodium falciparum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.4.2.1 0.56
-
 5'-methylthioinosine mutant enzyme V66I/Y73I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 4.6
-
 5'-methylthioinosine mutant enzyme V73I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 4.7
-
 5'-methylthioinosine mutant enzyme V66I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 6.8
-
 5'-methylthioinosine mutant enzyme Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 62
-
 5'-methylthioinosine mutant enzyme V66I/V73I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 71
-
 5'-methylthioinosine mutant enzyme V73I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 83
-
 Inosine mutant enzyme Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 90
-
 Inosine mutant enzyme V66I/Y73I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 94
-
 5'-methylthioinosine mutant enzyme V66I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 95
-
 5'-methylthioinosine wild type enzyme, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 130
-
 Inosine mutant enzyme V66I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 140
-
 Inosine mutant enzyme V73I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 150
-
 Inosine wild type enzyme, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 160
-
 Inosine mutant enzyme V73I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 170
-
 Inosine mutant enzyme V66I/V73I, at pH 7.4, temperature not specified in the publication Plasmodium falciparum
2.4.2.1 190
-
 Inosine mutant enzyme V66I/Y160F, at pH 7.4, temperature not specified in the publication Plasmodium falciparum