Literature summary for 2.7.13.3 extracted from

Cheung, J.; Hendrickson, W.A.
Crystal structures of C4-dicarboxylate ligand complexes with sensor domains of histidine kinases DcuS and DctB (2008), J. Biol. Chem., 283, 30256-30265.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of selenomethionine-labeled DctB residues 28-286 in Escherichia coli strain BL21(DE3)	Vibrio cholerae serotype O1
expression of selenomethionine-labeled DcuS residues 42-181 in Escherichia coli strain BL21(DE3) as GST-tagged protein	Escherichia coli K-12

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant selenomethionine-labeled and wild-type GST-tagged DcuS residues 42-181 in complex with C4-dicarboxylate ligand, 18.5 mg/ml protein, hanging-drop vapor diffusion, using a reservoir buffer containing 20-24% PEG monoethyl ether 2000, 15% isopropyl alcohol, 0.2 M ammonium citrate, and 0.1 M sodium acetate, pH 4.5, at 4 °C, 1 week, X-ray diffraction structure determination and analysis	Escherichia coli K-12
recombinant selenomethionine-labeled DctB residues 28-286 in complex with C4-dicarboxylate ligand, by hanging-drop vapor diffusion at 4°C against a buffer containing 6% isopropyl alcohol, 0.2 M calcium acetate, and 0.1 M Tris-HCl, pH 7.5, the protein concentration is 8.5 mg/ml with a protein to buffer ratio of 1:1, 2 weeks, X-ray diffraction structure determination and analysis	Vibrio cholerae serotype O1

Crystallization (Comment)

Organism

purified recombinant selenomethionine-labeled and wild-type GST-tagged DcuS residues 42-181 in complex with C4-dicarboxylate ligand, 18.5 mg/ml protein, hanging-drop vapor diffusion, using a reservoir buffer containing 20-24% PEG monoethyl ether 2000, 15% isopropyl alcohol, 0.2 M ammonium citrate, and 0.1 M sodium acetate, pH 4.5, at 4 °C, 1 week, X-ray diffraction structure determination and analysis

Escherichia coli K-12

recombinant selenomethionine-labeled DctB residues 28-286 in complex with C4-dicarboxylate ligand, by hanging-drop vapor diffusion at 4°C against a buffer containing 6% isopropyl alcohol, 0.2 M calcium acetate, and 0.1 M Tris-HCl, pH 7.5, the protein concentration is 8.5 mg/ml with a protein to buffer ratio of 1:1, 2 weeks, X-ray diffraction structure determination and analysis

Vibrio cholerae serotype O1

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
cell membrane	-	Vibrio cholerae serotype O1	-	-
cell membrane	-	Escherichia coli K-12	-	-
additional information	periplasmic sensor domains	Vibrio cholerae serotype O1	-	-
additional information	periplasmic sensor domains	Escherichia coli K-12	-	-

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
additional information	Escherichia coli K-12	a chemical or other stimulus is detected by the periplasmic sensor domain of a transmembrane histidine kinase sensor, which in turn relays a signal through a phosphotransfer cascade to the cognate cytoplasmic response regulator. Such systems lead ultimately to changes in gene expression or cell motility. Mechanisms of ligand binding and signal transduction through the cell membrane, overview	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli K-12	P0AEC8	gene dcuS	-
Vibrio cholerae serotype O1	-	gene dctB	-
Vibrio cholerae serotype O1 MO45	-	gene dctB	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant selenomethionine-labeled DctB residues 28-286 from Escherichia coli strain BL21(DE3) by anion exchange chromatography and gel filtration, followed by a second step of anion exchange chromatography	Vibrio cholerae serotype O1
recombinant selenomethionine-labeled GST-tagged DcuS residues 42-181 from Escherichia coli strain BL21(DE3) by glutathione affinity chromatography and gel filtration	Escherichia coli K-12

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	a chemical or other stimulus is detected by the periplasmic sensor domain of a transmembrane histidine kinase sensor, which in turn relays a signal through a phosphotransfer cascade to the cognate cytoplasmic response regulator. Such systems lead ultimately to changes in gene expression or cell motility. Mechanisms of ligand binding and signal transduction through the cell membrane, overview	Escherichia coli K-12	?	-	?

Subunits

Subunits	Comment	Organism
More	DctB shows a mixed alpha/beta-structure containing two subdomains of similar folds, each consisting of a five-stranded antiparallel beta-sheet flanked by helices on either side, structure analysis, residues 28-286 comprise the sensory domain of the enzyme, overview	Vibrio cholerae serotype O1
More	DcuS shows a mixed alpha/beta-structure containing two subdomains of similar folds, each consisting of a five-stranded antiparallel beta-sheet flanked by helices on either side, structure analysis, residues 42-181 comprise the sensory domain of the enzyme, dimerization of DcuS-42181, overview	Escherichia coli K-12

Synonyms

Synonyms	Comment	Organism
DctB	-	Vibrio cholerae serotype O1
DcuS	-	Escherichia coli K-12