Literature summary for 2.7.13.3 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.

Cloned(Commentary)

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

Crystallization (Commentary)

Crystallization (Comment)	Organism
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

Crystallization (Comment)

Organism

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

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

KM Value [mM]

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

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
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	-
additional information	the enzyme has a C-terminal cytoplasmic catalytic core of about 220 residues	Bacillus subtilis	-	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Bacillus subtilis

Natural Substrates/ Products (Substrates)

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

Organism

Organism	UniProt	Comment	Textmining
Bacillus subtilis	O34757	-	-
Bacillus subtilis 168	O34757	-	-

Purification (Commentary)

Purification (Comment)	Organism
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)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ATP + protein L-histidine	-	Bacillus subtilis	ADP + protein N-phospho-L-histidine	-	?
ATP + protein L-histidine	-	Bacillus subtilis 168	ADP + protein N-phospho-L-histidine	-	?
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	?	-	?
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

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

Synonyms

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

Temperature Optimum [°C]

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

pH Optimum

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

Cofactor

Cofactor	Comment	Organism	Structure
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

General Information	Comment	Organism
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
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
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