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Literature summary for 2.7.12.2 extracted from

  • Low, H.; Chua, C.S.; Sim, T.-S.
    Plasmodium falciparum possesses a unique dual-specificity serine/threonine and tyrosine kinase, Pfnek3 (2012), Cell. Mol. Life Sci., 69, 1523-1535.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
additional information the enzyme is activated by phosphorylation through a specific mitogen-activated protein kinase kinase kinase Plasmodium falciparum

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of GST-tagged wild-type and mutant Pfnek3 enzymes in Escherichia coli strain BL21 Plasmodium falciparum

Protein Variants

Protein Variants Comment Organism
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
Y117D site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities Plasmodium falciparum
Y117E site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities Plasmodium falciparum
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
Y122D site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities Plasmodium falciparum
Y122E site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities Plasmodium falciparum
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
Y172D site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities Plasmodium falciparum
Y172E site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities Plasmodium falciparum
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
Y238D site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities Plasmodium falciparum
Y238E site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities Plasmodium falciparum
Y238F site-directed mutagenesis, inactive mutant Plasmodium falciparum
Y286F site-directed mutagenesis, inactive mutant Plasmodium falciparum
Y99F site-directed mutagenesis, the mutation leads to drastic reductions in both Pfnek3 tyrosine phosphorylation and catalytic activities Plasmodium falciparum

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Plasmodium falciparum
Mn2+ highly activating, more effective than Mg2+ Plasmodium falciparum
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

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + myelin basic protein Plasmodium falciparum Pfnek3 phosphorylates MBP on threonine, but not serine residues ADP + phosphorylated myelin basic protein
-
?
ATP + Pfmap2 Plasmodium falciparum the enzyme phosphorylates its potential in vivo Pfmap2 substrate largely on Thr290 ADP + phosphorylated Pfmap2
-
?

Organism

Organism UniProt Comment Textmining
Plasmodium falciparum
-
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
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

Purification (Commentary)

Purification (Comment) Organism
recombinant GST-tagged wild-type and mutant Pfnek3 enzymes from Escherichia coli strain BL21 by glutathione affinity chromatography Plasmodium falciparum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + myelin basic protein Pfnek3 phosphorylates MBP on threonine, but not serine residues Plasmodium falciparum ADP + phosphorylated myelin basic protein
-
?
ATP + Pfmap2 the enzyme phosphorylates its potential in vivo Pfmap2 substrate largely on Thr290 Plasmodium falciparum ADP + phosphorylated Pfmap2
-
?
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 ?
-
?

Synonyms

Synonyms Comment Organism
MAPKK
-
Plasmodium falciparum
Pfnek3
-
Plasmodium falciparum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Plasmodium falciparum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.2
-
assay at Plasmodium falciparum

Cofactor

Cofactor Comment Organism Structure
ATP
-
Plasmodium falciparum

General Information

General Information Comment Organism
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