EC Number | Activating Compound | Comment | Organism | Structure |
---|---|---|---|---|
2.7.12.2 | additional information | the enzyme is activated by phosphorylation through a specific mitogen-activated protein kinase kinase kinase | Plasmodium falciparum |
EC Number | Cloned (Comment) | Organism |
---|---|---|
2.7.12.2 | recombinant expression of GST-tagged wild-type and mutant Pfnek3 enzymes in Escherichia coli strain BL21 | Plasmodium falciparum |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
2.7.12.2 | additional information | construction of a catalytically inactive enzyme knockout mutant DELTAPfnek3. The truncated Pfnek3 gene, encoding a catalytically active form of Pfnek3, is cloned into the GST-encoding pGEX-6P-1 vector | Plasmodium falciparum |
2.7.12.2 | Y117D | site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities | Plasmodium falciparum |
2.7.12.2 | Y117E | site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities | Plasmodium falciparum |
2.7.12.2 | Y117F | site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities. Reduction in tyrosine autophosphorylation is concomitant with the decrease in kinase activities for the Y117F, Y122F, and Y172F mutants | Plasmodium falciparum |
2.7.12.2 | Y122D | site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities | Plasmodium falciparum |
2.7.12.2 | Y122E | site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities | Plasmodium falciparum |
2.7.12.2 | Y122F | site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities. Reduction in tyrosine autophosphorylation is concomitant with the decrease in kinase activities for the Y117F, Y122F, and Y172F mutants | Plasmodium falciparum |
2.7.12.2 | Y172D | site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities | Plasmodium falciparum |
2.7.12.2 | Y172E | site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities | Plasmodium falciparum |
2.7.12.2 | Y172F | site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities. Reduction in tyrosine autophosphorylation is concomitant with the decrease in kinase activities for the Y117F, Y122F, and Y172F mutants | Plasmodium falciparum |
2.7.12.2 | Y238D | site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities | Plasmodium falciparum |
2.7.12.2 | Y238E | site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities | Plasmodium falciparum |
2.7.12.2 | Y238F | site-directed mutagenesis, inactive mutant | Plasmodium falciparum |
2.7.12.2 | Y286F | site-directed mutagenesis, inactive mutant | Plasmodium falciparum |
2.7.12.2 | Y99F | site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities | Plasmodium falciparum |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
2.7.12.2 | Mg2+ | required | Plasmodium falciparum | |
2.7.12.2 | Mn2+ | highly activating, more effective than Mg2+ | Plasmodium falciparum | |
2.7.12.2 | additional information | serine/threonine and tyrosine kinase activities are distinctly influenced by Mg2+ and Mn2+ cofactors. Pfnek3 can undergo further tyrosine autophosphorylation in vitro, with Mn2+ as the preferred metal cofactor | Plasmodium falciparum |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.7.12.2 | ATP + myelin basic protein | Plasmodium falciparum | Pfnek3 phosphorylates MBP on threonine, but not serine residues | ADP + phosphorylated myelin basic protein | - |
? | |
2.7.12.2 | ATP + Pfmap2 | Plasmodium falciparum | the enzyme phosphorylates its potential in vivo Pfmap2 substrate largely on Thr290 | ADP + phosphorylated Pfmap2 | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
2.7.12.2 | Plasmodium falciparum | - |
- |
- |
EC Number | Posttranslational Modification | Comment | Organism |
---|---|---|---|
2.7.12.2 | phosphoprotein | the enzyme is activated by tyroaisne phosphorylation through a specific mitogen-activated protein kinase kinase kinase. Tyrosine residues Y117, Y122, Y172, and Y238 are proposed as phosphorylation sites essential for mediating the catalytic activities of Pfnek3 | Plasmodium falciparum |
EC Number | Purification (Comment) | Organism |
---|---|---|
2.7.12.2 | recombinant GST-tagged wild-type and mutant Pfnek3 enzymes from Escherichia coli strain BL21 by glutathione affinity chromatography | Plasmodium falciparum |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.7.12.2 | ATP + myelin basic protein | Pfnek3 phosphorylates MBP on threonine, but not serine residues | Plasmodium falciparum | ADP + phosphorylated myelin basic protein | - |
? | |
2.7.12.2 | ATP + Pfmap2 | the enzyme phosphorylates its potential in vivo Pfmap2 substrate largely on Thr290 | Plasmodium falciparum | ADP + phosphorylated Pfmap2 | - |
? | |
2.7.12.2 | additional information | the enzyme displays both serine/threonine and tyrosine kinase activities in autophosphorylation reactions as well as in phosphorylation of the exogenous myelin basic protein substrate. Ability of Pfnek3 to autophosphorylate on both the serine/threonine and the tyrosine residues. The dual-specificity activity of the kinase is distinctly influenced by the type of divalent cation present | Plasmodium falciparum | ? | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
2.7.12.2 | MAPKK | - |
Plasmodium falciparum |
2.7.12.2 | Pfnek3 | - |
Plasmodium falciparum |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
2.7.12.2 | 30 | - |
assay at | Plasmodium falciparum |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
2.7.12.2 | 7.2 | - |
assay at | Plasmodium falciparum |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
2.7.12.2 | ATP | - |
Plasmodium falciparum |
EC Number | General Information | Comment | Organism |
---|---|---|---|
2.7.12.2 | additional information | Pfnek3 is a novel dual-specificity kinase of the malarial parasite, displaying both serine/threonine and tyrosine kinase activities, even though it has a HGDLKSTN motif in the catalytic loop that resembles the consensus HRDLKxxN signature found in the serine/threonine kinases. Tyrosine phosphorylation is involved in regulation of the serine/threonine and tyrosine kinase activities of Pfnek3 | Plasmodium falciparum |