Crystallization (Comment) | Organism |
---|---|
purified myristylated wild-type PKA and a K7C mutant as binary complex with bound substrate SP20 peptide and as ternary complex with bound substrate SP20 peptide and adenosine-5'-(beta,gamma-imido)triphosphate, hanging drop vapor diffusion method, 8-10 mg/ml protein in 50 mM N,N-bis(2-hydroxyethyl)glycine, 150 mM ammonium acetate, and 10 mM DTT, pH 8.0, is combined in 1:10:20:5 molar ratio of protein:AMP-PNP:Mg2+:SP20 for the ternary complex or 1:5 protein:SP20 for the binary complex, screening and method optimization, mixing of equal volumes of protein and well solution, the latter containing 2-18% 2-methyl-2,4-pentanediol, and mother liquor or buffers ranging from pH 5.35 to pH 8.5, 100 mM 2-[bis(2-hydroxyethyl)amino]-2-(hydroxymethyl) propane-1,3-diol, pH 6.5, and 9% MeOH, 8-10 weeks, 4°C, X-ray diffraction structure determination and analysis at 1.35-2.0 A resolution, modeling | Mus musculus |
Protein Variants | Comment | Organism |
---|---|---|
K7C | site-directed mutagenesis, the mutant exhibits altered kinetics in a myristylated state compared to the wild-type enzyme | Mus musculus |
N2D | site-directed mutagenesis, the mutant cannot be myristoylated at Asn2 | Mus musculus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.019 | - |
ATP | pH 7.0, temperature not specified in the publication, wild-type enzyme | Mus musculus | |
0.023 | - |
Leu-Arg-Arg-Ala-Ser-Leu-Gly | pH 7.0, temperature not specified in the publication, myristoylated wild-type enzyme | Mus musculus | |
0.023 | - |
ATP | pH 7.0, temperature not specified in the publication, myristoylated wild-type enzyme and enzyme mutant K7C | Mus musculus | |
0.027 | - |
Leu-Arg-Arg-Ala-Ser-Leu-Gly | pH 7.0, temperature not specified in the publication, enzyme mutant K7C | Mus musculus | |
0.029 | - |
Leu-Arg-Arg-Ala-Ser-Leu-Gly | pH 7.0, temperature not specified in the publication, wild-type enzyme | Mus musculus | |
0.032 | - |
ATP | pH 7.0, temperature not specified in the publication, myristoylated enzyme mutant K7C | Mus musculus | |
0.043 | - |
Leu-Arg-Arg-Ala-Ser-Leu-Gly | pH 7.0, temperature not specified in the publication, myristoylated enzyme mutant K7C | Mus musculus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Mus musculus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mus musculus | - |
- |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
lipoprotein | irreversible covalent N-myristylation of PKA or occupancy of the myristyl binding pocket may serve as a site for allosteric regulation in the catalytic C-subunit. And N-myristylation enhances the thermal stability of the enzyme, the myristylated C-subunit has a higher affinity for membranes alone. PKA cannot be myristylated if Asn2 is mutated to Asp | Mus musculus |
additional information | the C-subunit of PKA may be regulated by irreversible deamidation of Asn2. With PKA that is purified from tissues, irreversible deamidation of Asn2 to Asp or isoAsp occurs in about 1/3 of the total C-subunit protein. Also, the deamidated form of the protein has a higher cytosolic-to-nuclear ratio than the non-deamidated protein | Mus musculus |
phosphoprotein | phosphorylation on Ser10, Ser139, Thr197, and Ser338 | Mus musculus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + Leu-Arg-Arg-Ala-Ser-Leu-Gly | commercial artificial heptapeptide substrate kemptide | Mus musculus | ADP + Leu-Arg-Arg-Ala-phospho-Ser-Leu-Gly | - |
? | |
ATP + SP20 peptide | - |
Mus musculus | ADP + phosphorylated SP20 peptide | - |
? |
Synonyms | Comment | Organism |
---|---|---|
PKA | - |
Mus musculus |
protein kinase A | - |
Mus musculus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
18 | - |
Leu-Arg-Arg-Ala-Ser-Leu-Gly | pH 7.0, temperature not specified in the publication, myristoylated wild-type enzyme and enzyme mutant K7C | Mus musculus | |
19 | - |
Leu-Arg-Arg-Ala-Ser-Leu-Gly | pH 7.0, temperature not specified in the publication, wild-type enzyme | Mus musculus | |
29 | - |
Leu-Arg-Arg-Ala-Ser-Leu-Gly | pH 7.0, temperature not specified in the publication, myristoylated enzyme mutant K7C | Mus musculus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Mus musculus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Mus musculus |
General Information | Comment | Organism |
---|---|---|
metabolism | the catalytic subunit of cAMP-dependent protein kinase is a major target of cAMP signaling, and its regulation is of fundamental importance to biological processes | Mus musculus |