Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | activation loop structure and conformation, WNK1 performs autophosphorylation at S382 for activation | Rattus norvegicus |
Cloned (Comment) | Organism |
---|---|
expression of His-tagged low activity WNK1 kinase domain mutant S382A, comprising residues 194-483, in Escherichia coli | Rattus norvegicus |
Crystallization (Comment) | Organism |
---|---|
purified recombinant low activity WNK1 mutant S382A, wild-type and selenomethionine-labeled, hanging drop vapour diffusion method, 16°C, mixing of 0.002 ml of protein solution and of well solution, the latter containing 24% PEG monomethyl ester 2000, 0.3 M NaCl, 0.1 M HEPES, pH 7.0-8.0, 3 days, stabilization with 15% glycerol, X-ray diffraction structure determination and analysis at 1.8 A resolution | Rattus norvegicus |
Protein Variants | Comment | Organism |
---|---|---|
E268Q | site-directed mutagenesis, residue is probably important in enzyme stabilization since the mutant enzyme cannot be expressed recombinantly | Rattus norvegicus |
S382A | site-directed mutagenesis, mutant remains in the low activity form | Rattus norvegicus |
V318E/A448K | site-directed mutagenesis, the mutant shows 50-60% reduced activity compared to the wild-type WNK1 | Rattus norvegicus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Rattus norvegicus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Rattus norvegicus | WNK1 causes a hereditary form of hypertension, pseudohypoaldosteronism type II | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | - |
- |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
phosphoprotein | WNK1 performs autophosphorylation at S382 for activation | Rattus norvegicus |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged low activity WNK1 kinase domain mutant S382A, comprising residues 194-483, wild-type and selenomethionine-labeled, from Escherichia coli by chelating affinity and ion exchange chromatography, and gel filtration | Rattus norvegicus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ATP + a [protein]-(L-serine/L-threonine) = ADP + a [protein]-(L-serine/L-threonine) phosphate | active site and activation loop structure and conformation, Lys233, Cys250, Asp368, and Thr386 are important for activity, the substrate binding groove determines the substrate specificity via residues Val318 and Ala448 | Rattus norvegicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + synaptotagmin 2 | WNK1 specific substrate | Rattus norvegicus | ADP + phosphorylated synaptotagmin 2 | - |
? | |
additional information | WNK1 causes a hereditary form of hypertension, pseudohypoaldosteronism type II | Rattus norvegicus | ? | - |
? | |
additional information | WNK1 performs autophosphorylation at S382 for activation | Rattus norvegicus | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | kinase domain structure of WNK1 | Rattus norvegicus |
Synonyms | Comment | Organism |
---|---|---|
WNK1 | - |
Rattus norvegicus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.6 | - |
assay at | Rattus norvegicus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | binding site structure involving Lys233 | Rattus norvegicus |