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Literature summary extracted from

  • Trajtenberg, F.; Grana, M.; Ruetalo, N.; Botti, H.; Buschiazzo, A.
    Structural and enzymatic insights into the ATP binding and autophosphorylation mechanism of a sensor histidine kinase (2010), J. Biol. Chem., 285, 24892-24903.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.13.3 gene desK, recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain M15/pREP4 Bacillus subtilis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.13.3 purified recombinant DesK mutant H188V catalytic and ATP-binding domain in complex with ATP, by hanging drop vapour diffusion method, mixing of 0.002 ml of 10 mg/ml protein in 50 mM Tris-HCl, pH 8.0, 300 mM NaCl, 0.5 mM DTT, 10 mM MgCl2, 5 mM ATP, and 5 mM BeF3, with 0.002 ml of reservoir solution containing 20% PEG 3350, 0.2 M NH4ClX-ray diffraction structure determination and analysis at 1.8 A resolution Bacillus subtilis

Protein Variants

EC Number Protein Variants Comment Organism
2.7.13.3 E342A site-directed mutagenesis of the catalytic domain residue Bacillus subtilis
2.7.13.3 E343A site-directed mutagenesis of the catalytic domain residue Bacillus subtilis
2.7.13.3 G192C/G334C site-directed mutagenesis, the Cys-engineered mutant is used for interdomain disulfide covalent bonding studies Bacillus subtilis
2.7.13.3 H188V site-directed mutagenesis of the catalytic domain residue Bacillus subtilis
2.7.13.3 Q193C/G334C site-directed mutagenesis, the Cys-engineered mutant is used for interdomain disulfide covalent bonding studies Bacillus subtilis
2.7.13.3 S196C/G334C site-directed mutagenesis, the Cys-engineered mutant is used for interdomain disulfide covalent bonding studies Bacillus subtilis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.13.3 additional information
-
 additional information Michaelis-Menten kinetics Bacillus subtilis

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.7.13.3 membrane the enzyme structure displays an N-terminal sensor domain (about 150 residues) with almost no extracellular region, other than the loops that connect the four or five transmembrane segments Bacillus subtilis 16020
-
2.7.13.3 additional information the enzyme has a C-terminal cytoplasmic catalytic core of about 220 residues Bacillus subtilis
-
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.13.3 Mg2+ required Bacillus subtilis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.13.3 ATP + protein L-histidine Bacillus subtilis
-
 ADP + protein N-phospho-L-histidine
-
 ?
2.7.13.3 ATP + protein L-histidine Bacillus subtilis 168
-
 ADP + protein N-phospho-L-histidine
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.13.3 Bacillus subtilis O34757
-
-
2.7.13.3 Bacillus subtilis 168 O34757
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.13.3 recombinant N-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli strain M15/pREP4 by nickel affinity chromatography, dialysis, and gel filtration Bacillus subtilis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.13.3 ATP + protein L-histidine
-
 Bacillus subtilis ADP + protein N-phospho-L-histidine
-
 ?
2.7.13.3 ATP + protein L-histidine
-
 Bacillus subtilis 168 ADP + protein N-phospho-L-histidine
-
 ?
2.7.13.3 additional information the enzyme performs catalytic autophosphorylation, mechanism and kinetics, overview. DesK displays a compact structure at the ATP-binding pocket: the ATP lid loop is short with no secondary structural organization and becomes ordered upon ATP loading. Sequence conservation mapping onto the molecular surface, semi-flexible protein-protein docking simulations, and structure-based point mutagenesis present a specific domain-domain geometry during autophosphorylation catalysis. In vitro, DesKC catalyzes three different reactions depending on the phosphorylation states of the partners: its own phosphorylation, phosphotransfer to DesR, and dephosphorylation of phospho-DesR. Protein-protein docking and modelling of the enzyme in autophosphorylation state, residues involved in domain-domain interaction modulate catalysis, overview Bacillus subtilis ?
-
 ?
2.7.13.3 additional information the enzyme performs catalytic autophosphorylation, mechanism and kinetics, overview. DesK displays a compact structure at the ATP-binding pocket: the ATP lid loop is short with no secondary structural organization and becomes ordered upon ATP loading. Sequence conservation mapping onto the molecular surface, semi-flexible protein-protein docking simulations, and structure-based point mutagenesis present a specific domain-domain geometry during autophosphorylation catalysis. In vitro, DesKC catalyzes three different reactions depending on the phosphorylation states of the partners: its own phosphorylation, phosphotransfer to DesR, and dephosphorylation of phospho-DesR. Protein-protein docking and modelling of the enzyme in autophosphorylation state, residues involved in domain-domain interaction modulate catalysis, overview Bacillus subtilis 168 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.13.3 More analysis of the structure of enzyme catalytic and ATP-binding domain using the crystal structure, overview Bacillus subtilis

Synonyms

EC Number Synonyms Comment Organism
2.7.13.3 BSU19190
-
 Bacillus subtilis
2.7.13.3 class I HK
-
 Bacillus subtilis
2.7.13.3 DesK
-
 Bacillus subtilis
2.7.13.3 sensor histidine kinase
-
 Bacillus subtilis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.13.3 24
-
 assay at Bacillus subtilis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.13.3 7.6
-
 assay at Bacillus subtilis

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.13.3 ATP DesK displays a compact structure at the ATP-binding pocket: the ATP lid loop is short with no secondary structural organization and becomes ordered upon ATP loading. Sequence conservation mapping onto the molecular surface, semi-flexible protein-protein docking simulations, and structure-based point mutagenesis present a specific domain-domain geometry during autophosphorylation catalysis Bacillus subtilis

General Information

EC Number General Information Comment Organism
2.7.13.3 evolution the class I enzyme DesK belongs to the HK family HPK7, which includes the nitrogen metabolism regulators NarX/Q and the antibiotic sensor LiaS among other important sensor kinases Bacillus subtilis
2.7.13.3 additional information protein-protein docking analysis of the catalytic domain with the dimerization DHp domain of DesK, the C-terminal part of the ATP lid interacts with helix alpha1 of the DHp, through hydrogen bonds between His335 and Asp289 as well as Gly199 with Lys333, overview Bacillus subtilis
2.7.13.3 physiological function DesK is a sensor histidine kinase that allows Bacillus subtilis to respond to cold shock, triggering the adaptation of membrane fluidity via transcriptional control of a fatty acid desaturase. The transmembrane region can sense temperature-modulated fluidity changes of lipid bilayers, transmitting the signal toward the C-terminal cytoplasmic catalytic core of about 220 residues. The cold thermal stimulus is detected by DesK, which then interacts with its cognate response regulator, DesR, constituting a canonical two-component system, TCS Bacillus subtilis