Literature summary extracted from
Crawley, S.W.; Gharaei, M.S.; Ye, Q.; Yang, Y.; Raveh, B.; London, N.; Schueler-Furman, O.; Jia, Z.; Cote, G.P.
Autophosphorylation activates Dictyostelium myosin II heavy chain kinase A by providing a ligand for an allosteric binding site in the alpha-kinase domain (2011), J. Biol. Chem., 286, 2607-2616.
Activating Compound
EC Number |
Activating Compound |
Comment |
Organism |
Structure |
---|
2.7.11.7 |
additional information |
autophosphorylation at Thr825 activates MHCK A at least 50fold, dephosphorylation of the residue reduces the activity |
Dictyostelium discoideum |
|
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
2.7.11.7 |
expression of FLAG-tagged T825A and T825E enzyme mutants in Dictyostelium discoideum cells, expression of His-tagged wild-type and mutant MHCK A alpha-kinase domains in Escherichia coli strain BL21(DE3) |
Dictyostelium discoideum |
Crystallization (Commentary)
EC Number |
Crystallization (Comment) |
Organism |
---|
2.7.11.7 |
purified recombinant wild-type and truncation mutant enzymes, hanging drop vapor diffusion method, mixing of 0.001 ml protein solution with 0.001 ml of reservoir solution containing 0.1 M Tris-HCl, pH 8.5, 0.2 M NaH2PO4, and 18% w/v PEG 8000, 4°C, 10 days, X-ray diffraction structure determination and analysis at 1.9 A resolution |
Dictyostelium discoideum |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
2.7.11.7 |
additional information |
generation of a DELTA809 truncation mutant, phosphothreonine and the QQG(p)TMVMPD peptide restore the phosphorylation and also the ATPase activity of the mutant alpha-kinade domain, but mutations R734A and D762A abolish the ability of phosphothreonine to activate mutant alpha-kinase domain DELTA809 |
Dictyostelium discoideum |
2.7.11.7 |
Q822R/Q823R/T825S |
site-directed mutagenesis |
Dictyostelium discoideum |
2.7.11.7 |
T825A |
site-directed mutagenesis |
Dictyostelium discoideum |
2.7.11.7 |
T825E |
site-directed mutagenesis |
Dictyostelium discoideum |
2.7.11.7 |
T825S |
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme |
Dictyostelium discoideum |
Metals/Ions
EC Number |
Metals/Ions |
Comment |
Organism |
Structure |
---|
2.7.11.7 |
Mg2+ |
required, the Mg2 binding sites are regulatory, because millimolar concentrations of Mg2+, in excess of that required to bind to ATP, are required for A-CAT to exhibit maximal catalytic activity |
Dictyostelium discoideum |
|
Natural Substrates/ Products (Substrates)
EC Number |
Natural Substrates |
Organism |
Comment (Nat. Sub.) |
Natural Products |
Comment (Nat. Pro.) |
Rev. |
Reac. |
---|
2.7.11.7 |
ATP + [myosin heavy-chain] |
Dictyostelium discoideum |
- |
ADP + [myosin heavy-chain] phosphate |
- |
? |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
2.7.11.7 |
Dictyostelium discoideum |
P42527 |
- |
- |
Posttranslational Modification
EC Number |
Posttranslational Modification |
Comment |
Organism |
---|
2.7.11.7 |
phosphoprotein |
autophosphorylation at Thr825 activates MHCK A at least 50fold, dephosphorylation of the residue reduces the activity. The phosphorylated Thr825 docks intramolecularly into the phosphate-pocket. Allosteric activation is predicted to involve a conformational change in Arg734, which bridges the bound phosphate to Asp762 in a key active site loop |
Dictyostelium discoideum |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
2.7.11.7 |
recombinant His-tagged wild-type and mutant MHCK A alpha-kinase domains from Escherichia coli strain BL21(DE3) by nickel affinity chromatography |
Dictyostelium discoideum |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
2.7.11.7 |
ATP + [myosin heavy-chain] |
- |
Dictyostelium discoideum |
ADP + [myosin heavy-chain] phosphate |
- |
? |
|
2.7.11.7 |
additional information |
the enzyme performs autophosphorylation at Thr825 |
Dictyostelium discoideum |
? |
- |
? |
|
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
2.7.11.7 |
MHCK A |
- |
Dictyostelium discoideum |
2.7.11.7 |
myosin II heavy chain kinase A |
- |
Dictyostelium discoideum |
Temperature Optimum [°C]
EC Number |
Temperature Optimum [°C] |
Temperature Optimum Maximum [°C] |
Comment |
Organism |
---|
2.7.11.7 |
22 |
- |
assay at |
Dictyostelium discoideum |
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
2.7.11.7 |
7 |
- |
assay at |
Dictyostelium discoideum |
Cofactor
EC Number |
Cofactor |
Comment |
Organism |
Structure |
---|
2.7.11.7 |
ATP |
- |
Dictyostelium discoideum |
|
General Information
EC Number |
General Information |
Comment |
Organism |
---|
2.7.11.7 |
evolution |
myosin II heavy chain kinase A is a member of the atypical alpha-kinase family |
Dictyostelium discoideum |
2.7.11.7 |
malfunction |
the catalytic activity of A-CAT, the alpha-kinase domain of MHCKA comprising residues 552-841, is severely inhibited by the removal of a disordered C-terminal tail residues 806-841 |
Dictyostelium discoideum |
2.7.11.7 |
additional information |
the phosphate-pocket functions as a positive allosteric binding site, the phosphorylated Thr825 activates alpha-kinase domain by providing a covalently tethered ligand for the phosphate-pocket, mechanism modeling |
Dictyostelium discoideum |
2.7.11.7 |
physiological function |
the enzyme phosphorylates sites in the myosin II tail that block filament assembly |
Dictyostelium discoideum |