Literature summary for 4.2.1.10 extracted from

Light, S.H.; Minasov, G.; Shuvalova, L.; Peterson, S.N.; Caffrey, M.; Anderson, W.F.; Lavie, A.
A conserved surface loop in type I dehydroquinate dehydratases positions an active site arginine and functions in substrate binding (2011), Biochemistry, 50, 2357-2363.

Application

Application	Comment	Organism
drug development	the absence of DHQD in humans and its essentiality in many pathogenic bacteria make the enzyme a target for the development of nontoxic antimicrobials and design of type I DHQD inhibiting molecules	Salmonella enterica

Cloned(Commentary)

Cloned (Comment)	Organism
overexpression of His-tagged DHQD from MCSG7 expression vector in Escherichia coli strain BL21(DE3)	Salmonella enterica

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant DHQD by sitting drop vaour diffusion method, generation of three different crystal forms, the protein solution contains 500 mM NaCl and 10 mM Tris-HCl, pH 8.3, mixing of protein and reservoir solution in a 1:1 ratio at room temperature, the reservoir solutions contain: 200 mM MgCl2 and 20% PEG 3350, or 170 mM NH4OAc, pH 4.6, 25.5% PEG 4000, and 15% glycerol, or 200 mM MgCl2, 100 mM Tris, pH 8.5, and 20% PEG 3350 for the wild-type enzyme in open or in closed conformation of for the mutant Q236A, X-ray diffraction structure determination and analysis at 1.03-1.93 A resolution. The Q236A enzyme crystallized in another crystal form that diffracts to a significantly higher resolution than any of the wild-type crystals	Salmonella enterica

Crystallization (Comment)

Organism

purified recombinant DHQD by sitting drop vaour diffusion method, generation of three different crystal forms, the protein solution contains 500 mM NaCl and 10 mM Tris-HCl, pH 8.3, mixing of protein and reservoir solution in a 1:1 ratio at room temperature, the reservoir solutions contain: 200 mM MgCl2 and 20% PEG 3350, or 170 mM NH4OAc, pH 4.6, 25.5% PEG 4000, and 15% glycerol, or 200 mM MgCl2, 100 mM Tris, pH 8.5, and 20% PEG 3350 for the wild-type enzyme in open or in closed conformation of for the mutant Q236A, X-ray diffraction structure determination and analysis at 1.03-1.93 A resolution. The Q236A enzyme crystallized in another crystal form that diffracts to a significantly higher resolution than any of the wild-type crystals

Salmonella enterica

Protein Variants

Protein Variants	Comment	Organism
Q236A	site-directed mutagenesis, substrate binding structure and kinetics compared to the wild-type enzyme, the mutant shows highly reduced catalytic activity. The loop mutant fails to induce a conformational change in Arg213	Salmonella enterica
S232A	site-directed mutagenesis, substrate binding structure and kinetics compared to the wild-type enzyme, the mutant shows highly reduced catalytic activity	Salmonella enterica

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
3-dehydroquinate	Salmonella enterica	-	3-dehydroshikimate + H2O	-	?

Organism

Organism	UniProt	Comment	Textmining
no activity in Homo sapiens	-	-	-
Salmonella enterica	P58687	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged DHQD from Escherichia coli strain BL21(DE3) by nickel affinity chromatography	Salmonella enterica

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
3-dehydroquinate	-	Salmonella enterica	3-dehydroshikimate + H2O	-	?
additional information	functionality of a surface loop that closes over the active site following substrate binding, both direct and indirect mechanisms of involvement of the loop in substrate binding exist, overview. To establish a direct interaction with the substrate, closure of the loop necessitates a conformational change of a key active site arginine, which in turn positions the substrate productively. Loop closure induces a conformational change in Arg213. Specifically the side chain of Gln236, may be critical for inducing the change in the conformation of Arg213	Salmonella enterica	?	-	?

Synonyms

Synonyms	Comment	Organism
dehydroquinate dehydratase	-	Salmonella enterica
DHQD	-	Salmonella enterica
type I dehydroquinate dehydratase	-	Salmonella enterica
type I DHQD	-	Salmonella enterica

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
6.9	-	3-dehydroquinate	pH 8.3, temperature not specified in the publication, mutant Q236A	Salmonella enterica
107	-	3-dehydroquinate	pH 8.3, temperature not specified in the publication, mutant S232A	Salmonella enterica
210	-	3-dehydroquinate	pH 8.3, temperature not specified in the publication, wild-type enzyme	Salmonella enterica

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8.3	-	assay at	Salmonella enterica