Information on EC 2.7.6.3 - 2-amino-4-hydroxy-6-hydroxymethyldihydropteridine diphosphokinase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY
2.7.6.3
-
RECOMMENDED NAME
GeneOntology No.
2-amino-4-hydroxy-6-hydroxymethyldihydropteridine diphosphokinase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
ATP + 2-amino-4-hydroxy-6-hydroxymethyl-7,8-dihydropteridine = AMP + (2-amino-4-hydroxy-7,8-dihydropteridin-6-yl)methyl diphosphate
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
diphosphate transfer
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
6-hydroxymethyl-dihydropterin diphosphate biosynthesis I
-
6-hydroxymethyl-dihydropterin diphosphate biosynthesis II (archaea)
-
Folate biosynthesis
-
Metabolic pathways
-
SYSTEMATIC NAME
IUBMB Comments
ATP:2-amino-4-hydroxy-6-hydroxymethyl-7,8-dihydropteridine 6'-diphosphotransferase
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2-amino-4-hydroxy-6-hydroxymethyldihydropteridine pyrophosphokinase
-
-
-
-
6-hydroxy-7,8-dihydropterin pyrophosphokinase
-
-
-
-
6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase
-
-
6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase
P26281
-
6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase
-
-
6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase
-
-
6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase
-
-
6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase
-
-
6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase
Q7CKD7
-
6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase (HPPK)
-
-
6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase-dihydropteroate synthase
Q2A2W3
HPPK-DHPS, bifunctional enzyme
6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase-dihydropteroate synthase
Francisella tularensis LVS
Q2A2W3
HPPK-DHPS, bifunctional enzyme
-
6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase/dihydropteroate synthase
-
bifunctional enzyme
6-hydroxymethyl-7,8-dihydroxypterin pyrophosphokinase/7,8-dihydropteroate synthase
-
bifunctional protein involved in tetrahydrofolate synthesis
6-hydroxymethylpterin pyrophosphokinase
-
-
7,8-dihydro-6-hydroxymethylpterin pyrophosphokinase
-
-
-
-
7,8-dihydropteroate-synthesizing enzyme
-
it is not possible to distinguish, whether the enzyme exists as an enzyme complex of 6-hydroxymethyl-7,8-dihydroxypterin pyrophosphokinase and 7,8-dihydropteroate synthase or whether both reactions are catalyzed by a single enzyme
7,8-dihydroxymethylpterin-pyrophosphokinase
-
-
-
-
ATP:2-amino-4-hydroxy-6-hydroxymethyl-7,8-dihydropteridine 6'-pyrophosphotransferase
-
-
-
-
cytHPPK/DHPS
Q1ENB6
-
dihydropterin pyrophosphokinase
Q27865
-
H2-pteridine-CH2-OH pyrophosphokinase
-
-
-
-
H2-pteridine-CH2OH pyrophosphokinase
-
-
-
-
HPPK
-
-
-
-
HPPK
Francisella tularensis LVS
Q2A2W3
-
-
HPPK
Q7CKD7
-
HPPK/DHPS
-
-
HPPK/DHPS
-
bifunctional enzyme
HPPK/dihydropteroate synthase
-
HPPK part of the bifunctional protein can function by itself but a larger part of the polypeptide is needed to ensure full functionality
hydroxymethyl-7,8-dihydropterin pyrophosphokinase/7,8-dihydropteroate synthase
Q1ENB6
-
hydroxymethyldihydropteridine pyrophosphokinase
-
-
-
-
hydroxymethyldihydropterin diphosphokinase/dihydropteroate synthase
-
HPPK part of the bifunctional protein can function by itself but a larger part of the polypeptide is needed to ensure full functionality
hydroxymethyldihydropterin pyrophosphokinase (HPPK)
-
-
hydroxymethyldihydropterin pyrophosphokinase-dihydropteroate synthase
-
bifunctional enzyme
pyrophosphokinase, 2-amino-4-hydroxy-6-hydroxymethyldihydropteridine
-
-
-
-
mitHPPK/DHPS
Q1ENB6
-
additional information
-
6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase domain of the multifunctional folic acid synthesis enzyme
CAS REGISTRY NUMBER
COMMENTARY
37278-23-2
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
Francisella tularensis LVS
-
UniProt
Manually annotated by BRENDA team
Brookhaven Protein Data Bank: 1cbk
-
-
Manually annotated by BRENDA team
multifunctional folic acid synthesis fas gene of encodes dihydroneopterin aldolase, hydroxymethyldihydropterin pyrophosphokinase and dihydropteroate synthase
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
metabolism
-
6-hydroxymethyl-7,8-dihydropterin pyrophosphokinae is a committed enzyme in the folate pathway of Streptococcus aureus playing an integral role in the biosynthesis of tetrahydrofolate
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
6-hydroxymethyl-7,8-dihydropterin + ATP
6-hydroxymethyl-7,8-dihydropterin diphosphate + AMP
show the reaction diagram
Q27865
-
-
-
?
6-hydroxymethyl-7,8-dihydropterin + ATP
6-hydroxymethyl-7,8-dihydropterin diphosphate + AMP
show the reaction diagram
-
biosynthesis of folic acid
-
-
?
6-hydroxymethyl-7,8-dihydropterin + ATP
6-hydroxymethyl-7,8-dihydropterin diphosphate + AMP
show the reaction diagram
P53848
folate biosynthesis
-
-
?
6-hydroxymethyl-7,8-dihydropterin + ATP
6-hydroxymethyl-7,8-dihydropterin diphosphate + AMP
show the reaction diagram
-, Q1ENB6
tetrahydrofolate biosynthesis
-
-
?
6-hydroxymethyl-7,8-dihydropterin + MgATP2-
6-hydroxymethyl-7,8-dihydropterin diphosphate + MgAMP
show the reaction diagram
-
biosynthesis of folate cofactors
-
-
?
ATP + 2-amino-4-hydroxy-6-hydroxymethyl-7,8-dihydropteridine
AMP + 2-amino-7,8-dihydro-4-hydroxy-6-(diphosphooxymethyl)pteridine
show the reaction diagram
-
-
-
-
?
ATP + 2-amino-4-hydroxy-6-hydroxymethyl-7,8-dihydropteridine
AMP + 2-amino-7,8-dihydro-4-hydroxy-6-(diphosphooxymethyl)pteridine
show the reaction diagram
-
-
-
-
?
ATP + 2-amino-4-hydroxy-6-hydroxymethyl-7,8-dihydropteridine
AMP + 2-amino-7,8-dihydro-4-hydroxy-6-(diphosphooxymethyl)pteridine
show the reaction diagram
-
key step in biosynthesis of folic acid, ATP binding is followed by binding of 2-amino-4-hydroxy-6-hydroxymethyl-7,8-dihydropteridine
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropteridine
AMP + 6-hydroxymethyl-7,8-dihydropteridine diphosphate
show the reaction diagram
-
-
-
-
ir
ATP + 6-hydroxymethyl-7,8-dihydropteridine
AMP + 6-hydroxymethyl-7,8-dihydropteridine diphosphate
show the reaction diagram
-
-
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropteridine
AMP + 6-hydroxymethyl-7,8-dihydropteridine diphosphate
show the reaction diagram
-
-
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropteridine
AMP + 6-hydroxymethyl-7,8-dihydropteridine diphosphate
show the reaction diagram
-
-
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropteridine
AMP + 6-hydroxymethyl-7,8-dihydropteridine diphosphate
show the reaction diagram
-
-
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropteridine
AMP + 6-hydroxymethyl-7,8-dihydropteridine diphosphate
show the reaction diagram
-, Q2A2W3
-
-
-
ir
ATP + 6-hydroxymethyl-7,8-dihydropteridine
AMP + 6-hydroxymethyl-7,8-dihydropteridine diphosphate
show the reaction diagram
P26281
enzyme binds ATP first, followed by 2-amino-4-hydroxy-6-hydroxymethyl-7,8-dihydropteridine
-
?
ATP + 6-hydroxymethyl-7,8-dihydropteridine
AMP + 6-hydroxymethyl-7,8-dihydropteridine diphosphate
show the reaction diagram
-
ordered bi bi mechanism where ATP binds first and 6-hydroxymethyl-7,8-dihydropterin diphosphate is released last
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropteridine
AMP + 6-hydroxymethyl-7,8-dihydropteridine diphosphate
show the reaction diagram
P26281
the product 6-hydroxymethyl-7,8-dihydropterin diphosphate is an intermediate in the pathway for folic acid biosynthesis
-
?
ATP + 6-hydroxymethyl-7,8-dihydropteridine
AMP + 6-hydroxymethyl-7,8-dihydropteridine diphosphate
show the reaction diagram
-
ordered bi-bi mechanism with ATP as the first substrate
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropteridine
AMP + 6-hydroxymethyl-7,8-dihydropteridine diphosphate
show the reaction diagram
Francisella tularensis LVS
Q2A2W3
-
-
-
ir
ATP + 6-hydroxymethyl-7,8-dihydropteridine
AMP + 7,8-dihydro-6-(diphosphooxymethyl)pteridine
show the reaction diagram
-
-
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropterin
AMP + (7,8-dihydropterin-6-yl)methyl diphosphate
show the reaction diagram
P53848
-
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropterin
AMP + (7,8-dihydropterin-6-yl)methyl diphosphate
show the reaction diagram
-, Q1ENB6
-
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropterin
AMP + (7,8-dihydropterin-6-yl)methyl diphosphate
show the reaction diagram
Q7CKD7, -
-
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropterin
AMP + (7,8-dihydropterin-6-yl)methyl diphosphate
show the reaction diagram
-, Q1ENB6
6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase/7,8-dihydropteroate synthase, i.e. mitHPPK/DHPS, is a bifunctional mitochondrial enzyme, which catalyzes the first two consecutive steps of tetrahydrofolate biosynthesis, folate biosynthesis in Arabidopsis thaliana, overview
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropterin
AMP + (7,8-dihydropterin-6-yl)methyl diphosphate
show the reaction diagram
-
fourth step in folate biosynthesis, detailed overview
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropterin
AMP + (7,8-dihydropterin-6-yl)methyl diphosphate
show the reaction diagram
P53848
step in the folate biosynthetic pathway, overview
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropterin
AMP + (7,8-dihydropterin-6-yl)methyl diphosphate
show the reaction diagram
-
the enzyme is absolutely specific for the substrate
-
-
?
dATP + 2-amino-4-hydroxy-6-hydroxymethyl-7,8-dihydropteridine
dAMP + 2-amino-7,8-dihydro-4-hydroxy-6-(diphosphooxymethyl)pteridine
show the reaction diagram
-
-
-
-
?
additional information
?
-
P26281
binding affinity for GTP and GMP, 75fold weaker than for ATP
-
-
-
additional information
?
-
Q7CKD7, -
HPPK is a key enzyme in the folate-biosynthetic pathway and is essential for microorganisms but absent from mammals
-
-
-
additional information
?
-
P53848
the bifunctional HPPK-DHPS enzyme catalyzes sequential steps in the folate biosynthesis, i.e. the 6-hydroxymethylpterin pyrophosphokinase reaction and the dihydropteroate synthase, DHPS, reaction, EC 2.5.1.15, coupled assay method development using reduction of DHPS reaction product dihydropteroate to tetrahydropteroate by excess dihydrofolate reductase with the cofactor NADPH, the oxidation of NADPH is monitored, overview
-
-
-
additional information
?
-
-, Q2A2W3
the enzyme does not hydrolyze alpha,beta-methyleneadenosine 5-triphosphate
-
-
-
additional information
?
-
Francisella tularensis LVS
Q2A2W3
the enzyme does not hydrolyze alpha,beta-methyleneadenosine 5-triphosphate
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
6-hydroxymethyl-7,8-dihydropterin + ATP
6-hydroxymethyl-7,8-dihydropterin diphosphate + AMP
show the reaction diagram
Q27865
-
-
-
?
6-hydroxymethyl-7,8-dihydropterin + ATP
6-hydroxymethyl-7,8-dihydropterin diphosphate + AMP
show the reaction diagram
-
biosynthesis of folic acid
-
-
?
6-hydroxymethyl-7,8-dihydropterin + ATP
6-hydroxymethyl-7,8-dihydropterin diphosphate + AMP
show the reaction diagram
P53848
folate biosynthesis
-
-
?
6-hydroxymethyl-7,8-dihydropterin + ATP
6-hydroxymethyl-7,8-dihydropterin diphosphate + AMP
show the reaction diagram
-, Q1ENB6
tetrahydrofolate biosynthesis
-
-
?
6-hydroxymethyl-7,8-dihydropterin + MgATP2-
6-hydroxymethyl-7,8-dihydropterin diphosphate + MgAMP
show the reaction diagram
-
biosynthesis of folate cofactors
-
-
?
ATP + 2-amino-4-hydroxy-6-hydroxymethyl-7,8-dihydropteridine
AMP + 2-amino-7,8-dihydro-4-hydroxy-6-(diphosphooxymethyl)pteridine
show the reaction diagram
-
key step in biosynthesis of folic acid
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropteridine
AMP + 6-hydroxymethyl-7,8-dihydropteridine diphosphate
show the reaction diagram
-
-
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropteridine
AMP + 6-hydroxymethyl-7,8-dihydropteridine diphosphate
show the reaction diagram
P26281
the product 6-hydroxymethyl-7,8-dihydropterin diphosphate is an intermediate in the pathway for folic acid biosynthesis
-
?
ATP + 6-hydroxymethyl-7,8-dihydropterin
AMP + (7,8-dihydropterin-6-yl)methyl diphosphate
show the reaction diagram
Q7CKD7, -
-
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropterin
AMP + (7,8-dihydropterin-6-yl)methyl diphosphate
show the reaction diagram
-, Q1ENB6
6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase/7,8-dihydropteroate synthase, i.e. mitHPPK/DHPS, is a bifunctional mitochondrial enzyme, which catalyzes the first two consecutive steps of tetrahydrofolate biosynthesis, folate biosynthesis in Arabidopsis thaliana, overview
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropterin
AMP + (7,8-dihydropterin-6-yl)methyl diphosphate
show the reaction diagram
-
fourth step in folate biosynthesis, detailed overview
-
-
?
ATP + 6-hydroxymethyl-7,8-dihydropterin
AMP + (7,8-dihydropterin-6-yl)methyl diphosphate
show the reaction diagram
P53848
step in the folate biosynthetic pathway, overview
-
-
?
additional information
?
-
Q7CKD7, -
HPPK is a key enzyme in the folate-biosynthetic pathway and is essential for microorganisms but absent from mammals
-
-
-
additional information
?
-
P53848
the bifunctional HPPK-DHPS enzyme catalyzes sequential steps in the folate biosynthesis, i.e. the 6-hydroxymethylpterin pyrophosphokinase reaction and the dihydropteroate synthase, DHPS, reaction, EC 2.5.1.15
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Co2+
-
less effective than Mn2+ or Mg2+ in activation
KCl
-
activation at 0.4 M, inhibition at higher concentration
Mg2+
-
required for binding of nucleotides and for the binding of 6-hydroxymethyl-7,8-dihydropteridine
Mg2+
-
required, optimal activity at 4 mM
Mg2+
-
required
Mg2+
-, Q1ENB6
-
Mg2+
Q7CKD7, -
required
Mg2+
-
dependent on
Mg2+
-
required
Mg2+
-
required, 5 mM MgCl2 used in assay conditions
Mg2+
-, Q2A2W3
contains two Mg2+ ions
Mg2+
-
two Mg2+ ions are required for catalysis
Mn2+
-
can replace Mg2+ with a 10 mM optimum
NaCl
-
activation at 0.2 M, inhibition at higher concentration
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-Amino-4-hydroxy-6-carboxydihydropteridine
-
competitive with 2-amino-4-hydroxy-6-hydroxymethyldihydropteridine
2-amino-6-[(2-[4-[5-(6-amino-purin-9-yl)-3,4-dihydroxy-tetrahydrofuran-2-ylmethylsulfanyl]-piperidin-1-yl]-ethylamino)-methyl]-3H-pteridin-4-one
-
-
-
2-amino-6-[(2-[4-[5-(6-amino-purin-9-yl)-3,4-dihydroxy-tetrahydrofuran-2-ylmethylsulfanyl]-piperidin-1-yl]-ethylamino)-methyl]-7,7-dimethyl-7,8-dihydro-3H-pteridin-4-one
-
-
-
2-amino-7,7-dimethyl-4-oxo-3,4,7,8-tetrahydro-pteridine-6-carboxylic acid (2-[4-[5-(6-amino-purin-9-yl)-3,4-dihydroxy-tetrahydro-furan-2-ylmethylsulfanyl]-piperidin-1-yl]-ethyl)-amide
-
-
-
2-amino-7,7-dimethyl-4-oxo-3,4,7,8-tetrahydropteridine-6-carboxylic acid (2-[2-[5-(6-amino-purin-9-yl)-3,4-dihydroxy-tetrahydro-furan-2-ylmethanesulfonyl]-ethylcarbamoyl]-ethyl)-amide
-
about 45% residual activity at 0.01 mM, about 30% residual activity at 0.02 mM, about 15% residual activity at 0.05 mM, almost complete inhibition at 0.1 mM
-
5'-S-[1-(2-{[(2-amino-7,7-dimethyl-4-oxo-3,4,7,8-tetrahydropteridin-6-yl)carbonyl]amino}ethyl)piperidin-4-yl]-5'-thioadenosine
-
-
-
6-hydroxymethyl-7,7-dimethyl-7,8-dihydropterin
-
-
-
6-hydroxymethyl-7,8-dihydropterin diphosphate
-
competitive inhibitor of ATP, mixed type inhibitor of 6-hydroxymethyl-7,8-dihydropteridine
6-hydroxymethyl-7-methyl-7-phenethyl-7,8-dihydropterin
-
-
-
alpha,beta-methyleneadenosine triphosphate
-
competitive with respect to ATP
AMP
-
poor inhibitor
Guanidine-HCl
-
0.25 M, 50% inhibitioin
KCl
-
activation at 0.4 M, inhibition at higher concentration
NaCl
-
activation at 0.2 M, inhibition at higher concentration
P1-(6-hydroxymethylpterin)-P2-(5'-adenosyl)diphosphate
-
-
-
P1-(6-hydroxymethylpterin)-P3-(5'-adenosyl)triphosphate
-
-
-
P1-(6-hydroxymethylpterin)-P4-(5'-adenosyl)tetraphosphate
-
-
-
sulfachloropyridazine
P53848
-
sulfadoxine
-
competitive, moderately potent
sulfamethoxazole
P53848
-
sulfathiazole
-
competitive
Urea
-
0.9 M, 50% inhibition
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-, Q1ENB6
expression of cytHPPK/DHPS mRNA is induced by salt stress
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.01
-
2-amino-4-hydroxy-6-hydroxymethyldihydropteridine
-
pH 8.5, 37C
0.01
-
2-amino-4-hydroxy-6-hydroxymethyldihydropteridine
-
-
0.015
-
2-amino-4-hydroxy-6-hydroxymethyldihydropteridine
-
pH 8.6, 37C
0.015
-
2-amino-4-hydroxy-6-hydroxymethyldihydropteridine
-
-
0.001
-
6-hydroxymethyl-7,8-dihydropteridine
-
bifunctional enzyme 6-hydroxymethyl-7,8-dihydroxypterin pyrophosphokinase/7,8-dihydropteroate synthase
0.0016
-
6-hydroxymethyl-7,8-dihydropteridine
-
pH 8.0, 37C
0.002
-
6-hydroxymethyl-7,8-dihydropteridine
-
recombinant bifunctional enzyme 6-hydroxymethyl-7,8-dihydroxypterin pyrophosphokinase/7,8-dihydropteroate synthase expressed in Escherichia coli
0.0036
-
6-hydroxymethyl-7,8-dihydropteridine
-
pH 8.2, 50 mM Tris buffer
0.00039
-
6-hydroxymethyl-7,8-dihydropterin
P53848
-
0.011
-
ATP
-
pH 8.5, 37C
0.015
-
ATP
-
pH 8.6, 37C
0.015
-
ATP
-
-
0.017
-
ATP
-
pH 8.0, 37C
0.07
-
ATP
-
bifunctional enzyme 6-hydroxymethyl-7,8-dihydroxypterin pyrophosphokinase/7,8-dihydropteroate synthase from Pisum sativum and recombinant enzyme expressed in Escherichia coli
additional information
-
additional information
P53848
kinetics, HPPK-DHPS coupled assay method, overview
-
additional information
-
additional information
Q7CKD7, -
kinetic mechanism and thermodynamics, overview
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-
additional information
-
-
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.043
-
2-Amino-4-hydroxy-6-carboxydihydropteridine
-
pH 8.5, 37C
0.005
-
6-hydroxymethyl-7,8-dihydropterin diphosphate
-
versus ATP
0.013
-
6-hydroxymethyl-7,8-dihydropterin diphosphate
-
versus 6-hydroxymethyl-7,8-dihydropteridine
0.00031
-
alpha,beta-methyleneadenosine triphosphate
-
-
0.4
-
AMP
-
versus 6-hydroxymethyl-7,8-dihydropteridine
0.7
-
AMP
-
versus ATP
0.011
-
sulfadoxine
-
-
IC50 VALUE [mM]
IC50 VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.1
-
2-amino-6-[(2-[4-[5-(6-amino-purin-9-yl)-3,4-dihydroxy-tetrahydrofuran-2-ylmethylsulfanyl]-piperidin-1-yl]-ethylamino)-methyl]-3H-pteridin-4-one
-
IC50 above 0.1 mM, in 100 mM Tris, pH 8.3, temperature not specified in the publication
-
0.00316
-
2-amino-7,7-dimethyl-4-oxo-3,4,7,8-tetrahydro-pteridine-6-carboxylic acid (2-[4-[5-(6-amino-purin-9-yl)-3,4-dihydroxy-tetrahydro-furan-2-ylmethylsulfanyl]-piperidin-1-yl]-ethyl)-amide
-
in 100 mM Tris, pH 8.3, temperature not specified in the publication
-
0.00953
-
2-amino-7,7-dimethyl-4-oxo-3,4,7,8-tetrahydropteridine-6-carboxylic acid (2-[2-[5-(6-amino-purin-9-yl)-3,4-dihydroxy-tetrahydro-furan-2-ylmethanesulfonyl]-ethylcarbamoyl]-ethyl)-amide
-
in 100 mM Tris, pH 8.3, at 23C
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
-
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
8
-
-, Q1ENB6
assay at
8.3
-
Q7CKD7, -
assay at
8.5
9
-
Tris buffer
8.5
-
P53848
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.5
10.8
-
active between pH 7.5 and pH 10.8
8.5
10.5
-
pH 8.5: about 40% of maximal activity, pH 10.5: about 90% of maximal activity
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
-
-, Q1ENB6
assay at
37
-
P53848
assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
45
-
30C: about 40% of maximal activity, 37-45C: optimum
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.55
4.65
-
isoelectric focusing
7.1
-
-
calulation from amino acid sequence
9.1
-
-
isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-, Q1ENB6
developing, isozyme cytHPPK/DHPS gene is exclusively expressed in developing seeds, histochemical analysis of a transgenic cytHPPK/DHPS promoter-GUS line
Manually annotated by BRENDA team
additional information
-, Q1ENB6
ubiquitous expression of isozyme mitHPPK/DHPS
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-, Q1ENB6
isozyme cytHPPK/DHPS lacking a potential transit peptide
Manually annotated by BRENDA team
-, Q1ENB6
; mitHPPK/DHPS
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
Staphylococcus aureus (strain Mu50 / ATCC 700699)
Streptococcus pneumoniae (strain ATCC BAA-255 / R6)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20000
-
-
gel filtration, equilibrium sedimentation, quasi-elastic light scattering
25000
-
-
gel filtration
69000
-
-
calculation from multifunctional folic acid synthesis fas gene that encodes dihydroneopterin aldolase, hydroxymethyldihydropterin pyrophosphokinase and dihydropteroate synthase
150000
-
-
gel filtration
190000
-
-
gel filtration
330000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-, Q1ENB6
x * 54053, sequence calculation
?
-
x * 18000, SDS-PAGE
homodimer
Q7CKD7, -
Arg79 and Gln88 may help to stabilize the homodimer. The long side chain of Arg79 in YpHPPK forms many hydrophobic interactions with Trp90 and Gln49 from the partner subunit and a weak hydrogen bond with the side chain of Gln49, which may help to stabilize the dimeric molecule
monomer
-
1 * 25000, SDS-PAGE
monomer
-
1 * 18299, calculation from nucleotide sequence
monomer
-, Q2A2W3
1 * 50509, the bifunctional enzyme HPPK-DHPS exists mainly as a monomer in solution, calculated from amino acid sequence; 1 * 53000, the bifunctional enzyme HPPK-DHPS exists mainly as a monomer in solution, sedimentation velocity and equilibrium analysis
tetramer
-
4 * 83000, SDS-PAGE
monomer
Francisella tularensis LVS
-
1 * 50509, the bifunctional enzyme HPPK-DHPS exists mainly as a monomer in solution, calculated from amino acid sequence; 1 * 53000, the bifunctional enzyme HPPK-DHPS exists mainly as a monomer in solution, sedimentation velocity and equilibrium analysis
-
additional information
-
the bifunctional protein 6-hydroxymethyl-7,8-dihydroxypterin pyrophosphokinase/7,8-dihydropteroate synthase is involved in tetrahydrofolate synthesis
additional information
-
6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase domain of the multifunctional folic acid synthesis enzyme
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
apo W89A and its ternary complex with Mg-alpha,beta-methyleneadenosine triphosphate and 6-hydroxymethyl-7,8-dihydropterin are crystallized at 19C using the hanging-drop vapor-diffusion technique. The structure of the ternary complex is determined at 1.25 A resolution
-
complexed with inhibitor 2-amino-7,7-dimethyl-4-oxo-3,4,7,8-tetrahydropteridine-6-carboxylic acid (2-[2-[5-(6-amino-purin-9-yl)-3,4-dihydroxy-tetrahydro-furan-2-ylmethanesulfonyl]-ethylcarbamoyl]-ethyl)-amide, sitting drop vapor diffusion method, using 25% (w/v) PEG 3350 and 0.2 M NaCl in 0.1 M HEPES, pH 7.5
-
hanging-drop vapor-diffusion method. At 0.89-A resolution, two distinct conformations are observed for each of the two residues in the crystal structure of the wild-type enzyme in complex with two 6-hydroxymethyl-7,8-dihydropterin variants, two Mg2+ ions, and an ATP analogue. 1. Complex of wild-type enzyme with 6-hydroxymethylpterin, 6-carboxypterin and alpha,beta-methyleneadenosine 5'-triphosphate, 2. complex of mutant enzyme R82A with 6-hydroxymethyl-7,8-dihydropterin and alpha,beta-methyleneadenosine 5'-triphosphate, 3. complex of mutant enzyme R92A with 6-hydroxymethyl-7,8-dihydropterin and alpha,beta-methyleneadenosine 5'-triphosphate, 4. matant apoenzyme of R82A, 5. mutant apoenzyme of R92A, 6. mutant enzyme R92A in complex with Mg2+
-
in complex with 2-amino-6-[(2-{4-[5-(6-amino-purin-9-yl)-3,4-dihydroxy-tetrahydrofuran-2-ylmethylsulfanyl]-piperidin-1-yl}-ethylamino)-methyl]-3H-pteridin-4-one, 2-amino-6-[(2-{4-[5-(6-amino-purin-9-yl)-3,4-dihydroxy-tetrahydrofuran-2-ylmethylsulfanyl]-piperidin-1-yl}-ethylamino)-methyl]-7,7-dimethyl-7,8-dihydro-3H pteridin-4-one, or 2-amino-7,7-dimethyl-4-oxo-3,4,7,8-tetrahydro-pteridine-6-carboxylic acid (2-{4-[5-(6-amino-purin-9-yl)-3,4-dihydroxy-tetrahydro-furan-2-ylmethylsulfanyl]-piperidin-1-yl}-ethyl)-amide, sitting drop vapor diffusion method, using 20% or 25% (w/v) PEG 3350 as precipitant, at 19C
-
V83Gdel84-89 and its complex with alpha,beta--methyleneadenosine triphosphate and 6-hydroxymethyl-7,8-dihydropterin are crystallized at 19C using the hanging-drop vapor-diffusion technique
-
apoenzyme and in complex with substrate 6-hydroxymethyl-7,8-dihydropteridine or 2-(7-amino-1-methyl-4,5-dioxo-1,4,5,6-tetrahydorpyrimido[4,5-c]pyridazin-3-yl)propanoic acid, sitting drop vapor diffusion method, using 90 mM Tris (pH 8.0), 190 mM sodium acetate, 24% (w/v) polyethylene glycol (PEG) 4000, and 17% (v/v) glycerol, at 18C
-, Q2A2W3
a complex of the purified protein with a substrate analog is crystallized and its structure is solved by multiple anomalous dispersion using phase information obtained from a single crystal of selenomethionine-labeled protein
-
crystallization of a complex of the purified bifunctional polypeptide with a pterin monophosphate substrate analogue, structure solved by molecular replacement and refined to 2.3 A resolution. Three-dimensional structure in complex with the oxidized substrate analogue 6-hydroxy-methyl-pterin monophosphate reveals how the HPPK and DHPS functional domains associate at both the ternary and quaternary levels
-
sitting drop vapor diffusion method, using 1.08 M sodium malonate pH 7, 0.09 M bis-Tris pH 6.5, 0.175 M sodium formate, and 0.01 M sodium acetate pH 4.6
-
hanging-drop method
-
purified recombinant HPPK in complex with 6-hydroxymethyl-7,8-dihydropterin and an ATP analogue AMPCPP, 8.0 mg/ml HPPK in 15 mM HP, 25 mM AMPCPP, 50 mM MgCl2 and 10 mM Tris-HCl, pH 8.0 s mixed with well solution containing 0.18 M ammonium acetate, 30% w/v PEG 4000, 20 mM imidazole, and 0.1 M sodium acetate, pH 4.6, 1-3 weeks, X-ray diffraction structure determination and analysis, molecular replacement
Q7CKD7, -
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
55
-
-
complete inactivation
100
-
-
60 min, 75% loss of activity
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, enzyme loses activity over a period of months
-
-20C or -80C for 3 months in 20 mM Tris-HCl, pH 8.0 buffer containing 10 or 20% glycerol, no loss of activity. In buffer without glycerol, 30 and 50% of the enzyme activity are lost when stored for 6 months at -80C and -20C, respectively
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant N-terminally His6-tagged isozyme cytHPPK/DHPS from Escherichia coli strains DH-5alpha and BL-21AI
-, Q1ENB6
HPPK-glutathione S-transferase fusion proteins
-
HPPK-GST fusion proteins
-
Ni-NTA column chromatography and Superdex 75 gel filtration
-, Q2A2W3
recombinant bifunctional protein 6-hydroxymethyl-7,8-dihydroxypterin pyrophosphokinase/7,8-dihydropteroate synthase is involved in tetrahydrofolate
-
recombinant His6-tagged enzyme from Escherichia coli by metal affinity chromatography and ion exchange chromatography, cleavage of the tag by thrombin
P53848
Ni-NTA column chromatography and Superdex 75 gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli and Saccharomyces cerevisiae; isozyme cytHPPK/DHPS DNA and amino acid sequence determination and analysis, and phylogenetic analysis, transient expression of GFP-tagged isozyme cytHPPK/DHPS in the cytosol of Arabidopsis thaliana protoplasts using transfection via Agrobacterium tumefaciens strain GV3101/pMP90, functional complementation of a Saccharomyces cerevisiae mutant, that lacks the enzyme activity, by expression of isozyme cytHPPK/DHPS, expression and subcloning of N-terminally His6-tagged isozyme cytHPPK/DHPS in Escherichia coli strains DH-5alpha and BL-21AI, co-expression with GroESL chaperones
-, Q1ENB6
expressed in Escherichia coli BL21 cells
-, Q2A2W3
expression in Escherichia coli
-
bifunctional protein 6-hydroxymethyl-7,8-dihydroxypterin pyrophosphokinase/7,8-dihydropteroate synthase is involved in tetrahydrofolate expressed in Escherichia coli
-
expression in Escherichia coli
-
hydroxymethyldihydropterin diphosphokinase from Plasmodium falciparum complements a folK-knockout mutant in Escherichia coli when expressed as a separate polypeptide detached from dihydropteroate synthase. Hydroxymethyldihydropterin diphosphokinase part of the bifunctional protein can function by itself but that a larger part of the polypeptide is needed to ensure full functionality
-
multifunctional folic acid synthesis fas gene that encodes dihydroneopterin aldolase, hydroxymethyldihydropterin pyrophosphokinase and dihydropteroate synthase, in cultured Spodoptera frugiperda SF9 insect cells
-
the hydroxymethyldihydropterin pyrophosphokinase domain of the multifunctional folic acid synthesis Fas protein expressed as an independent enzyme in Escherichia coli, high level expression in inclusion bodies using an inducible tac promoter expression system
-
expression in Escherichia coli; expression of His6-tagged enzyme in Escherichia coli
P53848
expression of the bifunctional 6-hydroxymethyl-7,8-dihydropterin diphosphokinase/dihydropteroate synthase in Escherichia coli
-
expressed in Escherichia coli BL21(DE3) cells
-
expression in Escherichia coli
-
expression of His6-tagged maltose-binding-protein fusion HPPK in Escherichia coli strain BL21(DE3)
Q7CKD7, -
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
R82A
-
mutation causes a decrease in the rate constant for the chemical step by a factor of 380, no significant change in the binding energy or kinetics of either substrate
R84A
-
little changes in the dissociation constants and kinetic constants of the HPPK-catalyzed reaction
R84A
-
the mutation causes little changes in either dissociation constants or kinetic constants of the enzyme-catalyzed reaction except that the rate constant for the chemical step of the forward reaction decreases by a factor of 4
R92A
-
mutation causes a decrease in the rate constant for the chemical step by a factor of 35000. The mutation causes no significant change in the binding energy or binding kinetics of MgATP2-. It does not cause a significant change in the binding energy of 6-hydroxymethyl-7,8-dihydropterin either but causes a decrease in the association rate constant for the binding of 6-hydroxymethyl-7,8-dihydropterin by a factor of 1.4 and a decrease in the dissociation rate constant by a factor of 10
V83Gdel84-89
-
the deletion mutation does not have significant effects on the dissociation constants or the rate constants for the binding of the first substrate MgATP2- or its analogues. The dissociation constant of 6-hydroxymethyl-7,8-dihydropterin for the mutant increases by a factor of about 100, which is due to a large increase in the dissociation rate constant. The deletion mutation causes a shift of the rate-limiting step in the reaction and a decrease in the rate constant for the chemical step by a factor of 110000. The crystal structures reveal that the deletion mutation does not affect protein folding, but the catalytic center of the mutant is not fully assembled even upon the formation of the ternary complex and is not properly sealed. Loop 3 is dispensable for the folding of the protein and the binding of the first substrate MgATP2-, but is required for the assembling and sealing of the active center. The loop plays an important role in the stabilization of the ternary complex and is critical for catalysis
W89A
-
mutation increases the Kd for the binding of MgATP2- by a factor of 3, whereas the Kd for 6-hydroxymethyl-7,8-dihydropterin increases by a factor of 6, which is due to the increase in the dissociation rate constant. The mutation decreases the rate constant for the chemical step of the forward reaction by a factor of 15 and the rate constant for the chemical step of the reverse reaction by a factor of 25. The crystal structures of W89A show that W89A has different conformations in loops 2 and 3, but the critical catalytic residues are positioned for catalysis
W89A
-
the mutation does not have any significant effects on the Kd and the rate constants for the binding of MgATP, but the Kd for 6-hydroxymethyl-7,8-dihydropteridine of the mutant increases by a factor of 6.5. The mutation decreases the rate constant for the chemical step of the forward reaction by a factor of about 23 and the rate constant for the chemical step of the reverse reaction by a factor of about 33
additional information
-, Q1ENB6
construction of a T-DNA insertion mutant in the cytHPPK/DHPS gene, resulting in lower germination rates as compared with the wild-type upon application of oxidative and osmotic stress, phenotype, overview
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
medicine
-
because the enzyme is essential for microorganisms but is absent from human and animals, the enzyme is an excellent target for developing antimicrobial agent