Literature summary extracted from

Chiu, H.J.; Abdubek, P.; Astakhova, T.; Axelrod, H.L.; Carlton, D.; Clayton, T.; Das, D.; Deller, M.C.; Duan, L.; Feuerhelm, J.; Grant, J.C.; Grzechnik, A.; Han, G.W.; Jaroszewski, L.; Jin, K.K.; Klock, H.E.; Knuth, M.W.; Kozbial, P.; Krishna, S.S.; Kumar, A.; Marciano, D.; McMullan, D.; Miller, M.D.; Morse, A.T.
The structure of Haemophilus influenzae prephenate dehydrogenase suggests unique features of bifunctional TyrA enzymes (2010), Acta Crystallogr. Sect. F, 66, 1317-1325.

Cloned(Commentary)

EC Number	Cloned (Comment)	Organism
1.3.1.12	expression of His-tagged enzyme in Escherichia coli	Haemophilus influenzae

Crystallization (Commentary)

EC Number	Crystallization (Comment)	Organism
1.3.1.12	prephenate dehydrogenase component of the TyrA protein from strain Rd KW20 in complex with inhibitor tyrosine and cofactor NAD+, sitting drop vapour diffusion method, 200 nl of 19.6 mg/ml protein in 20 mM HEPES pH 8.0, 200 mM NaCl, 40 mM imidazole, 1 mM TCEP are mixed with 200 nl reservoir solution containing 0.04 M potassium dihydrogen phosphate, 20.0% v/v glycerol and 16.0% w/v PEG 8000, X-ray diffrraction structure determination and analysis at 2.0 A resolution	Haemophilus influenzae

Inhibitors

EC Number	Inhibitors	Comment	Organism	Structure
1.3.1.12	tyrosine	is bound directly at the catalytic site as a competitive inhibitor	Haemophilus influenzae

Natural Substrates/ Products (Substrates)

EC Number	Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
1.3.1.12	prephenate + NAD+	Haemophilus influenzae	-	4-hydroxyphenylpyruvate + CO2 + NADH + H+	-	?
1.3.1.12	prephenate + NAD+	Haemophilus influenzae KW20	-	4-hydroxyphenylpyruvate + CO2 + NADH + H+	-	?

Organism

EC Number	Organism	UniProt	Comment	Textmining
1.3.1.12	Haemophilus influenzae	P43902	-	-
1.3.1.12	Haemophilus influenzae KW20	P43902	-	-

Purification (Commentary)

EC Number	Purification (Comment)	Organism
1.3.1.12	recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography, cleavage of the tag by TEV protease	Haemophilus influenzae

Substrates and Products (Substrate)

EC Number	Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
1.3.1.12	additional information	a dimeric enzyme, with each monomer consisting of an N-terminal alpha/beta dinucleotide-binding domain and a C-terminal alpha-helical dimerization domain. Absence of an alpha/beta motif in HinfPDH that is present in other TyrA proteins. Residues from this motif are involved in discrimination between NADP+ and NAD+. The loop between beta5 and beta6 in the N-terminal domain is much shorter in HinfPDH and an extra helix is present at the C-terminus. Furthermore, HinfPDH adopts a more closed conformation compared with TyrA proteins that do not have tyrosine bound. This conformational change brings the substrate, cofactor and active-site residues into close proximity for catalysis. An ionic network consisting of Arg297, a key residue for tyrosine binding, a water molecule, Asp206, from the loop between beta5 and beta6, and Arg365', from the additional C-terminal helix of the adjacent monomer, is observed that might be involved in gating the active site. Active site structure, overview	Haemophilus influenzae	?	-	?
1.3.1.12	additional information	a dimeric enzyme, with each monomer consisting of an N-terminal alpha/beta dinucleotide-binding domain and a C-terminal alpha-helical dimerization domain. Absence of an alpha/beta motif in HinfPDH that is present in other TyrA proteins. Residues from this motif are involved in discrimination between NADP+ and NAD+. The loop between beta5 and beta6 in the N-terminal domain is much shorter in HinfPDH and an extra helix is present at the C-terminus. Furthermore, HinfPDH adopts a more closed conformation compared with TyrA proteins that do not have tyrosine bound. This conformational change brings the substrate, cofactor and active-site residues into close proximity for catalysis. An ionic network consisting of Arg297, a key residue for tyrosine binding, a water molecule, Asp206, from the loop between beta5 and beta6, and Arg365', from the additional C-terminal helix of the adjacent monomer, is observed that might be involved in gating the active site. Active site structure, overview	Haemophilus influenzae KW20	?	-	?
1.3.1.12	prephenate + NAD+	-	Haemophilus influenzae	4-hydroxyphenylpyruvate + CO2 + NADH + H+	-	?
1.3.1.12	prephenate + NAD+	key active-site residues are located at the domain interface, including His200, Arg297 and Ser179, that are involved in catalysis and/or ligand binding and are highly conserved in TyrA proteins	Haemophilus influenzae	4-hydroxyphenylpyruvate + CO2 + NADH + H+	-	?
1.3.1.12	prephenate + NAD+	-	Haemophilus influenzae KW20	4-hydroxyphenylpyruvate + CO2 + NADH + H+	-	?
1.3.1.12	prephenate + NAD+	key active-site residues are located at the domain interface, including His200, Arg297 and Ser179, that are involved in catalysis and/or ligand binding and are highly conserved in TyrA proteins	Haemophilus influenzae KW20	4-hydroxyphenylpyruvate + CO2 + NADH + H+	-	?

Subunits

EC Number	Subunits	Comment	Organism
1.3.1.12	dimer	SDS-PAGE and gel filtration, the dimeric enzyme, with each monomer consisting of an N-terminal alpha/beta dinucleotide-binding domain and a C-terminal alpha-helical dimerization domain, structure comparisons, overview	Haemophilus influenzae

Synonyms

EC Number	Synonyms	Comment	Organism
1.3.1.12	PDH	-	Haemophilus influenzae
1.3.1.12	tyrA	-	Haemophilus influenzae

Cofactor

EC Number	Cofactor	Comment	Organism	Structure
1.3.1.12	NAD+	binding mode	Haemophilus influenzae

General Information

EC Number	General Information	Comment	Organism
1.3.1.12	metabolism	chorismate mutase/prephenate dehydrogenase from Haemophilus influenzae Rd KW20 is a bifunctional enzyme that catalyzes the rearrangement of chorismate to prephenate and the NAD(P)+-dependent oxidative decarboxylation of prephenate to 4-hydroxyphenylpyruvate in tyrosine biosynthesis	Haemophilus influenzae