Literature summary for 2.7.4.22 extracted from

Meier, C.; Carter, L.G.; Sainsbury, S.; Mancini, E.J.; Owens, R.J.; Stuart, D.I.; Esnouf, R.M.
The crystal structure of UMP kinase from Bacillus anthracis (BA1797) reveals an allosteric nucleotide-binding site (2008), J. Mol. Biol., 381, 1098-1105.

Activating Compound

Activating Compound	Comment	Organism	Structure
GTP	UTP inhibition, but not GTP activation, is sensitive to Mg2+ levels	Bacillus anthracis

Application

Application	Comment	Organism
medicine	interest in new therapies against anthrax has arisen from its potential for bioterrorism. The allosteric pocket of the enzyme, with its atypical configuration of side chains, may provide a particularly suitable site for pharmacological intervention	Bacillus anthracis

Cloned(Commentary)

Cloned (Comment)	Organism
expression of the His-tagged enzyme in Escherichia coli strain B834(DE3)	Bacillus anthracis
vector pDEST14 is transformed into Escherichia coli B834(DE3)	Bacillus anthracis

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme in complex with ATP, 300 nl sitting drops containing 1.5 mg/mL purified protein are mixed with 3.3 mM ATP, 0.33 mM MgCl2, 66 mM Li2SO4, 3% PEG 3000, 33 mM imidazole, pH 8.0, and 20 mM spermine tetrahydrochloride, equilibration against a reservoir solution of 200 mM Li2SO4, 10% PEG 3000, and 100 mM imidazole, pH 8.0, cryoprotection in 70% reservoir solution with 30% glycerol, 2 days, X-ray diffraction structure determination and analysis at 2.82 A resolution	Bacillus anthracis
UMP kinase in complex with ATP and Mg2+ at 2.82 A resolution, 0.2 M lithium sulfate, 10% polyethylene glycol 3000, 0.1 M imidazole, pH 8.0, space group P 61 2 2, allosteric nucleotide-binding site identified, a structural model for the allosteric regulation presented	Bacillus anthracis

Crystallization (Comment)

Organism

purified recombinant enzyme in complex with ATP, 300 nl sitting drops containing 1.5 mg/mL purified protein are mixed with 3.3 mM ATP, 0.33 mM MgCl2, 66 mM Li2SO4, 3% PEG 3000, 33 mM imidazole, pH 8.0, and 20 mM spermine tetrahydrochloride, equilibration against a reservoir solution of 200 mM Li2SO4, 10% PEG 3000, and 100 mM imidazole, pH 8.0, cryoprotection in 70% reservoir solution with 30% glycerol, 2 days, X-ray diffraction structure determination and analysis at 2.82 A resolution

Bacillus anthracis

UMP kinase in complex with ATP and Mg2+ at 2.82 A resolution, 0.2 M lithium sulfate, 10% polyethylene glycol 3000, 0.1 M imidazole, pH 8.0, space group P 61 2 2, allosteric nucleotide-binding site identified, a structural model for the allosteric regulation presented

Bacillus anthracis

Inhibitors

Inhibitors	Comment	Organism	Structure
UTP	UTP inhibition, but not GTP activation, is sensitive to Mg2+ levels	Bacillus anthracis

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
cytosol	-	Bacillus anthracis	5829	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	-	Bacillus anthracis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
ATP + UMP	Bacillus anthracis	-	ADP + UDP	-	?
ATP + UMP	Bacillus anthracis	an essential metabolic step with a structure-based mechanism for the allosteric behavior of bacterial enzyme, overview	ADP + UDP	-	?
ATP + UMP	Bacillus anthracis BA1797	an essential metabolic step with a structure-based mechanism for the allosteric behavior of bacterial enzyme, overview	ADP + UDP	-	?
ATP + UMP	Bacillus anthracis BA1797	-	ADP + UDP	-	?

Organism

Organism	UniProt	Comment	Textmining
Bacillus anthracis	-	-	-
Bacillus anthracis	Q81S73	vector pDEST14 is transformed into Escherichia coli B834(DE3)	-
Bacillus anthracis BA1797	-	-	-
Bacillus anthracis BA1797	Q81S73	vector pDEST14 is transformed into Escherichia coli B834(DE3)	-

Purification (Commentary)

Purification (Comment)	Organism
Ni-NTA affinity chromatography and gel filtration	Bacillus anthracis
recombinant His-tagged enzyme from Escherichia coli strain B834(DE3) by nickel affinity chromatography and gel filtration	Bacillus anthracis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ATP + UMP	-	Bacillus anthracis	ADP + UDP	-	?
ATP + UMP	an essential metabolic step with a structure-based mechanism for the allosteric behavior of bacterial enzyme, overview	Bacillus anthracis	ADP + UDP	-	?
ATP + UMP	-	Bacillus anthracis BA1797	ADP + UDP	-	?
ATP + UMP	an essential metabolic step with a structure-based mechanism for the allosteric behavior of bacterial enzyme, overview	Bacillus anthracis BA1797	ADP + UDP	-	?

Subunits

Subunits	Comment	Organism
hexamer	trimer of dimers forming a staggered hexagonal ring	Bacillus anthracis
homohexamer	-	Bacillus anthracis

Synonyms

Synonyms	Comment	Organism
More	the enzyme belongs to the NMP family	Bacillus anthracis
UMP kinase	-	Bacillus anthracis
uridine monophosphate kinase	-	Bacillus anthracis
uridylate kinase	-	Bacillus anthracis

Cofactor

Cofactor	Comment	Organism	Structure
ATP	the cofactor, in addition to binding in the active sites, also interacts with separate binding pocket, with an allosteric binding site, located near the center of the hexameric structure	Bacillus anthracis