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Literature summary for 2.7.13.3 extracted from

  • Lee, J.; Tomchick, D.R.; Brautigam, C.A.; Machius, M.; Kort, R.; Hellingwerf, K.J.; Gardner, K.H.
    Changes at the KinA PAS-A dimerization interface influence histidine kinase function (2008), Biochemistry, 47, 4051-4064.
    View publication on PubMed

Cloned(Commentary)

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 (Commentary)

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

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

Metals/Ions Comment Organism Structure
Mg2+
-
Bacillus subtilis

Natural Substrates/ Products (Substrates)

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

Organism UniProt Comment Textmining
Bacillus subtilis P16497 strain 1A40
-
Bacillus subtilis 168 P16497 strain 1A40
-

Posttranslational Modification

Posttranslational Modification Comment Organism
phosphoprotein KinA performs autophosphorylation Bacillus subtilis

Specific Activity [micromol/min/mg]

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 and Products (Substrate)

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

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

Synonyms Comment Organism
KINA
-
Bacillus subtilis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Bacillus subtilis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Bacillus subtilis

Cofactor

Cofactor Comment Organism Structure
ATP
-
Bacillus subtilis