Literature summary for 2.7.1.189 extracted from

Ha, J.H.; Hauk, P.; Cho, K.; Eo, Y.; Ma, X.; Stephens, K.; Cha, S.; Jeong, M.; Suh, J.Y.; Sintim, H.O.; Bentley, W.E.; Ryu, K.S.
Evidence of link between quorum sensing and sugar metabolism in Escherichia coli revealed via cocrystal structures of LsrK and HPr (2018), Sci. Adv., 4, eaar7063 .

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Cloned(Commentary)

Cloned (Comment)	Organism
gene lsrK, sequence comparisons, recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)	Escherichia coli

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant LsrK/HPr complexes, i.e. selenomethionine-labeled His-tagged LsrK/HPr, His-tagged LsrK/HPr, His-tagged LsrK/HPr/ADP, and His-tagged LsrK/HPr/ATP, hanging drop vapor diffusion method, 15 mg/ml protein complex is mixed with reservoir buffer containing 15 to 20% v/v PEG 400, 0.1 M sodium phosphate-citrate, pH 4.2, and 0.2 M lithium sulfate, with or without addition of 5 mM MgCl2, and 5 mM ATP, or ADP, or ATPgammaS, X-ray diffraction structure determination and analysis at 2.7-3.2 A resolution	Escherichia coli

Crystallization (Comment)

Organism

purified recombinant LsrK/HPr complexes, i.e. selenomethionine-labeled His-tagged LsrK/HPr, His-tagged LsrK/HPr, His-tagged LsrK/HPr/ADP, and His-tagged LsrK/HPr/ATP, hanging drop vapor diffusion method, 15 mg/ml protein complex is mixed with reservoir buffer containing 15 to 20% v/v PEG 400, 0.1 M sodium phosphate-citrate, pH 4.2, and 0.2 M lithium sulfate, with or without addition of 5 mM MgCl2, and 5 mM ATP, or ADP, or ATPgammaS, X-ray diffraction structure determination and analysis at 2.7-3.2 A resolution

Escherichia coli

Inhibitors

Inhibitors	Comment	Organism
HPr	a phosphocarrier protein central to the sugar phosphotransferase system of Escherichia coli. HPr copurifies with LsrK. LsrK activity is inhibited when bound to HPr. HPr inhibited LsrK activity in a mixed manner of competitive and uncompetitive, not via competitive or uncompetitive inhibition alone. Phosphorylation of HPr reduces its inhibitory effect on LsrK	Escherichia coli
HPr H15A	the HPr H15A mutant displays almost no inhibition of the enzyme	Escherichia coli
HPr H15E	the HPr H15E mutant displays several times lower inhibition compared to the wild-type	Escherichia coli

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.058	-	(S)-4,5-dihydroxypentane-2,3-dione	recombinant enzyme, pH and temperature not specified in the publication	Escherichia coli

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
intracellular	-	Escherichia coli	5622	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + (S)-4,5-dihydroxypentane-2,3-dione	Escherichia coli	-	ADP + (S)-4-hydroxypentane-2,3-dione 5-phosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	P77432	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, anion exchange chromatography, and gel filtration	Escherichia coli

Source Tissue

Source Tissue	Comment	Organism	Textmining
additional information	HPr, a phosphocarrier protein central to the sugar phosphotransferase system of Escherichia coli, copurifies with LsrK. LsrK activity is increased in the stationary phase	Escherichia coli	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + (S)-4,5-dihydroxypentane-2,3-dione	-	Escherichia coli	ADP + (S)-4-hydroxypentane-2,3-dione 5-phosphate	-	?
ATP + HPr	-	Escherichia coli	ADP + phospho-HPr	-	?

Subunits

Subunits	Comment	Organism
monomer	HisLsrK-HPr protein complex, crystal structure and gel filtration	Escherichia coli

Synonyms

Synonyms	Comment	Organism
AI-2 kinase	UniProt	Escherichia coli
lsrK	-	Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
17	-	(S)-4,5-dihydroxypentane-2,3-dione	recombinant enzyme, pH and temperature not specified in the publication	Escherichia coli

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Escherichia coli

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.00008	-	HPr	inhibition of recombinant enzyme, pH and temperature not specified in the publication	Escherichia coli
0.0012	-	HPr H15E	inhibition of recombinant enzyme, pH and temperature not specified in the publication	Escherichia coli

General Information

General Information	Comment	Organism
metabolism	linkages between quorum sensing (QS) activity and sugar metabolism, model for LsrK/HPr interplay between QS and PTS, overview	Escherichia coli
physiological function	quorum sensing (QS), a bacterial process that regulates population-scale behavior, is mediated by small signaling molecules, called autoinducers (AIs), that are secreted and perceived, modulating a collective phenotype. Because the autoinducer AI-2 is secreted by a wide variety of bacterial species, its perception cues bacterial behavior. This response is mediated by the lsr (LuxS-regulated) operon that includes the AI-2 transporter LsrACDB and the kinase LsrK. The AI-2-mediated QS system is partially regulated by substrate availability and cell metabolism. LuxS synthesizes AI-2 as a by-product of the activated methyl cycle, after which AI-2 accumulates extracellularly. AI-2 is imported by LsrACDB and phosphorylated by the kinase LsrK, sequestering it within the cell. LsrK is tightly bound to the PTS protein HPr, and HPr directly influences LsrK activity	Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
293.1	-	(S)-4,5-dihydroxypentane-2,3-dione	recombinant enzyme, pH and temperature not specified in the publication	Escherichia coli