Cloned (Comment) | Organism |
---|---|
expression of residues 10-117 of KinA, fused to an N-terminal His6Gbeta1-tag and a TEV protease site in the tag-protein linker, in Escherichia coli strain BL21(DE3), expresssion of the selenomethionine variant in Escherichia coli strain B834 | Bacillus subtilis |
Crystallization (Comment) | Organism |
---|---|
purified recombinant residues 10-117 of KinA, fused to an N-terminal His6Gbeta1-tag and a TEV protease site in the tag-protein linker, free or as selenomethionine-tagged variant, hanging-drop vapor-diffusion method, 20°C, mixing of 0.001 ml 7 mg/mL protein solution containing 25 mM Tris pH 8.0, 100 mM NaCl, with 0.001 ml well solution containing 13-15% w/v PEG 10000, 0.1 M ammonium acetate, and 0.1 M bis-Tris, pH 5.5, and with 20 mM DTT in case of the selenomethionine-labeled protein, 4 days, cryoprotetion using 25% v/v glycerol, X-ray diffraction structure determination and analysis at 1.7-2.0 A resolution | Bacillus subtilis |
Protein Variants | Comment | Organism |
---|---|---|
I108A | the point mutation does not affect KinA autophosphorylation activity, but interfers with KinA PAS-A dimerization | Bacillus subtilis |
I95A | the point mutation slightly reduces the KinA autophosphorylation activity and interfers with KinA PAS-A dimerization | Bacillus subtilis |
I95E | the point mutation reduces the KinA autophosphorylation activity and interfers with KinA PAS-A dimerization | Bacillus subtilis |
additional information | a significant decrease occurs of the autophosphorylation rate of a KinA protein lacking the N-terminal Per-ARNT-Sim domain, which plays a critical role in the catalytic activity of this enzyme | Bacillus subtilis |
Y29A | the point mutation is an activating mutation in full-length KinA, but interfers with KinA PAS-A dimerization shifting the PAS-A monomer/dimer equilibrium significantly toward the monomeric form | Bacillus subtilis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Bacillus subtilis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Bacillus subtilis | the N-terminal Per-ARNT-Sim domain plays a critical role in the catalytic activity of this enzyme, a significant decrease occurs of the autophosphorylation rate of a KinA protein lacking this domain | ? | - |
? | |
additional information | Bacillus subtilis 168 | the N-terminal Per-ARNT-Sim domain plays a critical role in the catalytic activity of this enzyme, a significant decrease occurs of the autophosphorylation rate of a KinA protein lacking this domain | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | P16497 | strain 1A40 | - |
Bacillus subtilis 168 | P16497 | strain 1A40 | - |
Posttranslational Modification | Comment | Organism |
---|---|---|
phosphoprotein | KinA performs autophosphorylation | Bacillus subtilis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
1.3 | - |
KinA autophosphorylation, purified recombinant mutant I95A | Bacillus subtilis |
2.3 | - |
KinA autophosphorylation, purified recombinant wild-type enzyme | Bacillus subtilis |
2.8 | - |
KinA autophosphorylation, purified recombinant mutant I108A | Bacillus subtilis |
4 | - |
KinA autophosphorylation, purified recombinant mutant Y29A | Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | the N-terminal Per-ARNT-Sim domain plays a critical role in the catalytic activity of this enzyme, a significant decrease occurs of the autophosphorylation rate of a KinA protein lacking this domain | Bacillus subtilis | ? | - |
? | |
additional information | KinA performs autophosphorylation | Bacillus subtilis | ? | - |
? | |
additional information | the N-terminal Per-ARNT-Sim domain plays a critical role in the catalytic activity of this enzyme, a significant decrease occurs of the autophosphorylation rate of a KinA protein lacking this domain | Bacillus subtilis 168 | ? | - |
? | |
additional information | KinA performs autophosphorylation | Bacillus subtilis 168 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | wild-type KinA PAS-A, NMR, SDS-PAGE and gel filtration | Bacillus subtilis |
More | determination of the importance of several residues at the dimer interface for KinA enzymatic activity in vitro and in vivo, KinA architecture and domain structure, and PAS-A domain organization and secondary structure, overview | Bacillus subtilis |
Synonyms | Comment | Organism |
---|---|---|
KINA | - |
Bacillus subtilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Bacillus subtilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Bacillus subtilis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Bacillus subtilis |