Information on EC 2.5.1.63 - adenosyl-fluoride synthase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.5.1.63
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RECOMMENDED NAME
GeneOntology No.
adenosyl-fluoride synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
S-adenosyl-L-methionine + fluoride = 5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
adenosyl group transfer
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-
-
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halogenation
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-
-
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
fluoroacetate and fluorothreonine biosynthesis
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-
SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:fluoride adenosyltransferase
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CAS REGISTRY NUMBER
COMMENTARY hide
438583-16-5
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
gene flA
UniProt
Manually annotated by BRENDA team
gene flA
UniProt
Manually annotated by BRENDA team
solated in 2011 from Ghana, gene flA
UniProt
Manually annotated by BRENDA team
solated in 2011 from Ghana, gene flA
UniProt
Manually annotated by BRENDA team
gene flA4
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2'-deoxyadenosyl-L-methionine + fluoride
2'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
5'-deoxy-5'-fluoroadenosine + L-selenomethionine
Se-adenosyl-L-selenomethionine + fluoride
show the reaction diagram
-
-
-
-
r
methylaza-S-adenosyl-L-methionine + fluoride
?
show the reaction diagram
S-adenosyl-L-methionine + chloride
5'-deoxy-5'-chloroadenosine + L-methionine
show the reaction diagram
-
-
-
-
r
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
additional information
?
-
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fluoroacetate and 4-fluorothreonine accumulate in the fermentation media of Streptomyces cattleya at the mM level, when a fluoride source is added to the growth medium
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
K+
-
in the coupled assay with SAM synthase and fluorinase
Mn2+
slightly activating
additional information
-
Mg2+ at 1 mM does not affect enzyme activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
S-adenosyl-L-homocysteine
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competent, competitive
sinefungin
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weak
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.36 - 36.8
fluoride
0.0008 - 0.42
S-adenosyl-L-methionine
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000012 - 2
fluoride
0.000083 - 0.074
S-adenosyl-L-methionine
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.029
S-adenosyl-homocysteine
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-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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development of a coupled assay with SAM synthase and fluorinase
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.8
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assay at
7.9
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assay at
8.2
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recombinant enzyme with sbstrate methylaza-S-adenosyl-L-methionine
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 9.5
activity range, profile overview
5.5 - 10
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activity range, profile overview
6.5 - 9.5
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activity range, profile overview
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
32000
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6 * 32000, SDS-PAGE and electrospray mass spectrometry
34402
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6 * 34402, (alpha,beta)3, recombinant enzyme, ESI-MS mass spectrometry
180000
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gel filtration
200000
recombinant enzyme, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
additional information
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the N-terminal domain has a central seven-stranded beta-sheet, which combines parallel and antiparallel strands sandwiched between alpha-helices, the C-terminal domain is composed of a five- and a four-stranded antiparallel beta-sheet
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified enzyme in complex with 5',5'-difluorodeoxy-2-ethynyladenosine and tartrate, X-ray diffraction structure determination and analysis at 1.8 A resolution, molecular replacement using structure with PDB ID 1RQR
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purified enzyme in complex with 5'-fluorodeoxy-2-ethynyladenosine, X-ray diffraction structure determination and analysis at 2.4 A resolution, molecular replacement
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purified recombinant enzyme, free from tags or bound adenosine, complexed to 2'-deoxy-5'-fluoroadenosine, 4 mg/ml protein is incubated with 20 mM ligand at 25°C for 4 h, followed by crystallization using vapour diffusion against a reservoir containing 30% PEG 1000, 0.1 M phosphate-citrate pH 4.5, 0.2 M Li2SO4, X-ray diffraction structure determination and analysis at 2.4 A resolution, molecular replacement
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purified recombinant wild-type and selenomethionine labelled enzyme with bound S-adenosyl-L-methionine, X-ray diffraction structure determination and analysis at 1.9 A resolution, structure modelling of monomers a and b
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structure modelling of the fluoride ion/SAM prereaction complex at the active site of the fluorinase immediately prior to SN2 substitution, fluoride ion is modeled into the SAM-fluorinase cocomplex, which is solved by X-ray crystallography, overview
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GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
water stabilizes the enzyme, thus water-absorbing polymer and ionic liquid is used to immobilize the enzyme and keep it surrounded by water
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
nickel-affinity column
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recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography and gel filtration
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recombinant His-tagged enzyme from Escherichia coli strain BL21-Gold (DE3) pLysS by nickel affinity chromatography and gel filtration
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recombinant His-tagged enzyme from Escherichia coli strain BL21-Gold(DE3)pLysS by nickel affinity chromatography and gel filtration
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recombinant wild-type and mutant His6-tagged enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
recombinant wild-type His6-tagged enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli BL21; expression in Escherichia coli BL21(DE3)
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expression of His-tagged enzyme in Escherichia coli BL21(DE3) pLysS with plasmid pET28b+
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gene flA, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of wild-type His6-tagged enzymes in Escherichia coli strain BL21(DE3)
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gene flA, DNA sequence determination and analysis, overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3) and in strain B834(DE3), the latter results in the selenomethionine labeled enzyme
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gene flA, genetic structure and organization
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gene flA, genetic structure and organization, the flA gene is located in close proximity to the corresponding flB gene
gene flA, located at the center of the 10 kb gene Spencer cluster, genetic structure and organization, the f lA gene is located in close proximity to the corresponding f lB gene. Funtional recombinant expression in Salinispora tropica using gene replacement and replacing the gene salL that encodes chlorinase, EC 2.5.1.94, by gene flA from Streptomaces cattleya. The mutated Salinispora tropica strain produces fluorosalinosporamide
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gene flA, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21-Gold (DE3) pLysS
gene nobA, DNA and amino acid sequence determination and analysis, sequence comparisons, recombinant expression of wild-type and mutant His6-tagged enzymes in Escherichia coli strain BL21(DE3)
mutant enzymes expressed in Escherichia coli C43 (DE3) cells
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the fluorometabolite pathway is upregulated by increased fluoride ion levels, perhaps due to upregulation of f lG encoding a putative DNA binding regulatory protein
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D16A
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inactive
D16N
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3% activity compared to the wild type enzyme
D16S
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inactive
F156A
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3% activity compared to the wild type enzyme
F156V
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25% activity compared to the wild type enzyme
S158A
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38% activity compared to the wild type enzyme
S158G
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8% activity compared to the wild type enzyme
T80A
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15% activity compared to the wild type enzyme
T80S
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95% activity compared to the wild type enzyme
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
biotechnology
pharmacology
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a two-step radiolabelling protocol of a cancer relevant cRGD peptide is described where the fluorinase enzyme is used to catalyse a transhalogenation reaction to generate [18F]-5'-fluoro-5'-deoxy-2-ethynyladenosine, followed by a click reaction to an azide tethered cRGD peptide. This protocol offers efficient radiolabelling of a biologically relevant peptide construct in water at pH 7.8, 37°C in 2 hours, which is metabolically stable in rats and retains high affinity for alphavbeta3 integrin
synthesis