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Information on EC 2.7.7.2 - FAD synthase and Organism(s) Homo sapiens and UniProt Accession Q8NFF5

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EC Tree
     2 Transferases
         2.7 Transferring phosphorus-containing groups
             2.7.7 Nucleotidyltransferases
                2.7.7.2 FAD synthase
IUBMB Comments
Requires Mg2+ and is highly specific for ATP as phosphate donor . The cofactors FMN and FAD participate in numerous processes in all organisms, including mitochondrial electron transport, photosynthesis, fatty-acid oxidation, and metabolism of vitamin B6, vitamin B12 and folates . While monofunctional FAD synthetase is found in eukaryotes and in some prokaryotes, most prokaryotes have a bifunctional enzyme that exhibits both this activity and that of EC 2.7.1.26, riboflavin kinase [3,5].
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This record set is specific for:
Homo sapiens
UNIPROT: Q8NFF5
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Word Map
The taxonomic range for the selected organisms is: Homo sapiens
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
Synonyms
fads1, flad1, fad synthase, fad pyrophosphorylase, flavin adenine dinucleotide synthetase, atp:fmn adenylyltransferase, atribf1, atribf2, fmn:atp adenylyltransferase, mj1179, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ATP:FMN adenylyl transferase
-
FAD synthetase
-
FAD synthetase isoform 1
-
FAD synthetase isoform 2
-
FADS2
adenosine triphosphate-riboflavin mononucleotide transadenylase
-
-
-
-
adenosine triphosphate-riboflavine mononucleotide transadenylase
-
-
-
-
FAD pyrophosphorylase
-
-
-
-
FADS2
-
-
FMN adenylyltransferase
-
-
-
-
FMN pyrophosphorylase
-
-
-
-
lysZ
-
-
-
-
riboflavin adenine dinucleotide pyrophosphorylase
-
-
-
-
riboflavin mononucleotide adenylyltransferase
-
-
-
-
riboflavine adenine dinucleotide adenylyltransferase
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
nucleotidyl group transfer
-
-
-
-
PATHWAY SOURCE
PATHWAYS
-
-, -, -, -
SYSTEMATIC NAME
IUBMB Comments
ATP:FMN adenylyltransferase
Requires Mg2+ and is highly specific for ATP as phosphate donor [5]. The cofactors FMN and FAD participate in numerous processes in all organisms, including mitochondrial electron transport, photosynthesis, fatty-acid oxidation, and metabolism of vitamin B6, vitamin B12 and folates [3]. While monofunctional FAD synthetase is found in eukaryotes and in some prokaryotes, most prokaryotes have a bifunctional enzyme that exhibits both this activity and that of EC 2.7.1.26, riboflavin kinase [3,5].
CAS REGISTRY NUMBER
COMMENTARY hide
9026-37-3
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + FMN
diphosphate + FAD
show the reaction diagram
diphosphate + FAD
ATP + FMN
show the reaction diagram
ATP + FMN
diphosphate + FAD
show the reaction diagram
ATP + roseoflavin mononucleotide
diphosphate + roseoflavin adenine dinucleotide
show the reaction diagram
-
-
-
-
?
diphosphate + FAD
ATP + FMN
show the reaction diagram
-
-
-
-
r
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
FAD
ration of FAD/monomer is 0.6
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
Ca2+ gives maximum FAD synthesis equal to 62% of that measured with Mg2+
Co2+
in the presence of about 0.5 mM Co2+, the activity is almost equal to that measured with Mg2+
MgCl2
strictly required
Mn2+
Mn2+ gives maximum FAD synthesis equal to 77% of that measured with Mg2+
Co2+
-
Co2+ induces FAD hydrolysis, which is strongly stimulated in the presence of K+, K0.5 value is 0.035 mM
K+
-
stimulates Co2+-induced FAD hydrolysis, K0.5 value 7.2
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
GDP
competitive inhibition
GTP
uncompetitive inhibition
Urea
the enzymatic activity of isoform FADS2 decreases dramatically at increasing urea concentration, with a mid-point for activity loss at 2.1 m urea
KI
-
inhibits FAD hydrolysis
NADH
-
inhibits FAD hydrolysis
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
reduced glutathione
-
stimulates FAD hydrolysis
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00691 - 0.0153
ATP
0.042
diphosphate
pH 7.5, 37°C
0.00013 - 1.5
FMN
0.068 - 0.109
FMN
0.116
roseoflavin mononucleotide
-
with 24 mM Na2S2O4, in 50 mM potassium phosphate (pH 7.5), at 37°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00691
ATP
pH 7.5, 37°C
0.00013 - 0.078
FMN
0.02 - 0.08
FMN
0.07
roseoflavin mononucleotide
-
with 24 mM Na2S2O4, in 50 mM potassium phosphate (pH 7.5), at 37°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.83 - 1.2
FMN
0.6
roseoflavin mononucleotide
-
with 24 mM Na2S2O4, in 50 mM potassium phosphate (pH 7.5), at 37°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.5
GDP
at 37°C, in 50 mM Tris-HCl, pH 7.5
1.8
GTP
at 37°C, in 50 mM Tris-HCl, pH 7.5
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.03
at 37°C
0.59
crude cell extract, at 37°C
6.8
after 12.5fold purification, at 37°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5
-
hydrolysis of FAD
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.66
isoelectric focusing
6
calculated from amino acid sequence
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
FAD1_HUMAN
587
0
65266
Swiss-Prot
Mitochondrion (Reliability: 2)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
36500
gel filtration
54190
calculated from amino acid sequence
54200
x * 54200 Da, predicted
55000
SDS-PAGE, T7-tagged enzyme
55840
calculated from amino acid sequence, T7-tagged enzyme
55844
x * 55000, SDS-PAGE, x * 55844, calculated
56537
x * 56537, isoform FADS2, calculated from amino acid sequence
57000
x * 57000, isoform FADS2, SDS-PAGE
60000
SDS-PAGE, isoform 2
63000
SDS-PAGE
65270
calculated from amino acid sequence
65300
x * 65300 Da, predicted
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
1 * 38266, calculated from sequence, 1 * 36000, SDS-PAGE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
homology modeling of structure
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
51.6
Tm value
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Ni2+-chelating column chromatography and hydroxyapatite column chromatography
Ni2+-chelating Sepharose Fast Flow column chromatography
recombinant protein
Ni2+-nitrilotriacetic-agarose column chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
expressed in Escherichia coli strain Rosetta(DE3)
expressed in Escherichia coli, in BHK-21 cell, and Caco-2 cell
expression in Escherichia coli
His6-tagged enzyme is expressed in Escherichia coli Rosetta (DE3) cells
expressed in Escherichia coli strains BL21 and Rosetta (DE3) and in Pichia pastoris strain X33
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Brizio, C.; Galluccio, M.; Wait, R.; Torchetti, E.M.; Bafunno, V.; Accardi, R.; Gianazza, E.; Indiveri, C.; Barile, M.
Over-expression in Escherichia coli and characterization of two recombinant isoforms of human FAD synthetase
Biochem. Biophys. Res. Commun.
344
1008-1016
2006
Homo sapiens (Q8NFF5), Homo sapiens
Manually annotated by BRENDA team
Galluccio, M.; Brizio, C.; Torchetti, E.M.; Ferranti, P.; Gianazza, E.; Indiveri, C.; Barile, M.
Over-expression in Escherichia coli, purification and characterization of isoform 2 of human FAD synthetase
Protein Expr. Purif.
52
175-181
2007
Homo sapiens (Q8NFF5), Homo sapiens
Manually annotated by BRENDA team
Torchetti, E.M.; Brizio, C.; Colella, M.; Galluccio, M.; Giancaspero, T.A.; Indiveri, C.; Roberti, M.; Barile, M.
Mitochondrial localization of human FAD synthetase isoform 1
Mitochondrion
10
263-273
2010
Homo sapiens (Q8NFF5), Homo sapiens
Manually annotated by BRENDA team
Pedrolli, D.B.; Nakanishi, S.; Barile, M.; Mansurova, M.; Carmona, E.C.; Lux, A.; Gaertner, W.; Mack, M.
The antibiotics roseoflavin and 8-demethyl-8-amino-riboflavin from Streptomyces davawensis are metabolized by human flavokinase and human FAD synthetase
Biochem. Pharmacol.
82
1853-1859
2011
Homo sapiens
Manually annotated by BRENDA team
Torchetti, E.; Bonomi, F.; Galluccio, M.; Gianazza, E.; Giancaspero, T.; Iametti, S.; Indiveri, C.; Barile, M.
Human FAD synthase (isoform 2): A component of the machinery that delivers FAD to apo-flavoproteins
FEBS J.
278
4435-4449
2011
Homo sapiens (Q8NFF5)
-
Manually annotated by BRENDA team
Leone, P.; Galluccio, M.; Brizio, C.; Barbiroli, A.; Iametti, S.; Indiveri, C.; Barile, M.
The hidden side of the human FAD synthase 2
Int. J. Biol. Macromol.
138
986-995
2019
Homo sapiens
Manually annotated by BRENDA team
Leone, P.; Galluccio, M.; Barbiroli, A.; Eberini, I.; Tolomeo, M.; Vrenna, F.; Gianazza, E.; Iametti, S.; Bonomi, F.; Indiveri, C.; Barile, M.
Bacterial production, characterization and protein modeling of a novel monofuctional isoform of FAD synthase in humans An emergency protein?
Molecules
23
116
2018
Homo sapiens (Q8NFF5), Homo sapiens
Manually annotated by BRENDA team