Information on EC 2.1.1.107 - uroporphyrinogen-III C-methyltransferase

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

EC NUMBER
COMMENTARY
2.1.1.107
-
RECOMMENDED NAME
GeneOntology No.
uroporphyrinogen-III C-methyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
S-adenosyl-L-methionine + precorrin-1 = S-adenosyl-L-homocysteine + precorrin-2
show the reaction diagram
-
-
-
-
S-adenosyl-L-methionine + uroporphyrinogen III = S-adenosyl-L-homocysteine + precorrin-1
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
methyl group transfer
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
Metabolic pathways
-
Porphyrin and chlorophyll metabolism
-
SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:uroporphyrinogen-III C-methyltransferase
This enzyme catalyses two sequential methylation reactions, the first forming precorrin-1 and the second leading to the formation of precorrin-2. It is the first of three steps leading to the formation of siroheme from uroporphyrinogen III. The second step involves an NAD+-dependent dehydrogenation to form sirohydrochlorin from precorrin-2 (EC 1.3.1.76, precorrin-2 dehydrogenase) and the third step involves the chelation of Fe2+ to sirohydrochlorin to form siroheme (EC 4.99.1.4, sirohydrochlorin ferrochelatase). In Saccharomyces cerevisiae, the last two steps are carried out by a single bifunctional enzyme, Met8p. In some bacteria, steps 1-3 are catalysed by a single multifunctional protein called CysG, whereas in Bacillus megaterium, three separate enzymes carry out each of the steps, with SirA being responsible for the above reaction. Also involved in the biosynthesis of cobalamin.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
adenosylmethionine-uroporphyrinogen III methyltransferase
-
-
-
-
CobA/HemD
Lactobacillus reuteri CRL 1098
-
-
-
S-adenosyl-L-methionine dependent uroporphyrinogen III methylase
-
-
-
-
S-adenosyl-L-methionine-dependent uroporphyrinogen III methyltransferase
P95417
-
S-adenosyl-L-methionine: uroporphyrinogen III methyltransferase
-
-
SAM-dependent uroporphyrinogen III methyltransferase
-
-
sirohaem synthase
-
-
-
-
SUMT
Geobacillus stearothermophilus V
Q6TA16
-
-
SUMT
-
-
uroporphyrinogen III C-methyltransferase
-
-
-
-
uroporphyrinogen III methylase
-
-
-
-
uroporphyrinogen III methyltransferase
-
-
uroporphyrinogen III methyltransferase
-
-
uroporphyrinogen III methyltransferase/synthase
-
-
uroporphyrinogen III synthase/methyltransferase
-
-
uroporphyrinogen III synthase/methyltransferase
Lactobacillus reuteri CRL 1098
-
-
-
uroporphyrinogen methyltransferase,
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
73665-99-3
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
genomic DNA, cDNA has accession number L47479
SwissProt
Manually annotated by BRENDA team
gene cobA/hemD
-
-
Manually annotated by BRENDA team
Geobacillus stearothermophilus V
gene cobA
UniProt
Manually annotated by BRENDA team
Lactobacillus reuteri CRL 1098
-
-
-
Manually annotated by BRENDA team
strain PAO1, gene nirE
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
metabolism
-
the enzyme is involved in the tetrapyrrole biosynthesis. Uroporphyrinogen III represents the first branch point in the pathway, where the action of enzymes such as HemE, F (N) and G results in the formation of protoporphyrin IX, a precursor of modified tetrapyrroles such as haem and chlorophyll. Alternatively, uroporphyrinogen III can undergo a S-adenosylmethionine-dependent transmethylation positions 2 and 7, by CysG, Met1p or CobA depending on the organism, to generate precorrin-2, a highly unstable yellow dipyrrocorphin, overview
metabolism
-
the enzyme is important in the heme d1 biosynthesis, the essential prosthetic group of the dissimilatory nitrite reductase cytochrome cd1 requiring the methylation of the tetrapyrrole precursor uroporphyrinogen III at positions C-2 and C-7, pathway overview
metabolism
-
uroporphyrinogen III synthase/methyltransferase is a key bifunctional enzyme in the biosynthesis of cobalamin and other tetrapyrrols
metabolism
Lactobacillus reuteri CRL 1098
-
uroporphyrinogen III synthase/methyltransferase is a key bifunctional enzyme in the biosynthesis of cobalamin and other tetrapyrrols
-
physiological function
-, Q6TA16
the role of SUMT methyltransferase in tellurite(ate) detoxification is not related to tellurium volatilization
physiological function
Geobacillus stearothermophilus V
-
the role of SUMT methyltransferase in tellurite(ate) detoxification is not related to tellurium volatilization
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
-
produces equal amounts of precorrin-1 and precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-, Q51720
-
after prolonged incubation (15 h) also tri- and tetra-methylated compounds formed, third methyl group at position 12
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
-
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
-
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
-
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
-
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
-
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
P21632
-
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-, Q51720
-
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-, Q96532
-
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
methylates positions 2 and 7
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
methylates positions 2 and 7
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
methylates positions 2 and 7
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
methylates positions 2 and 7
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
involved in biosynthesis of sirohydrochlorin
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
involved in biosynthesis of sirohydrochlorin
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-, Q51720
involved in biosynthesis of sirohydrochlorin
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
involved in biosynthesis of siroheme
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
involved in biosynthesis of siroheme
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-, Q96532
involved in biosynthesis of siroheme
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
involved in biosynthesis of cobalamin
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
involved in biosynthesis of cobalamin
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
P21632
involved in biosynthesis of cobalamin
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-, Q51720
involved in biosynthesis of cobalamin
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-, Q96532
no synthesis of cobalamin
product is precorrin-2
?
S-adenosyl-L-methionine + heptacarboxyporphyrinogen III
?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + hexacarboxyporphyrinogen III
?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + pentacarboxyporphyrinogen III
?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + uroporphyrinogen I
?
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + uroporphyrinogen I
?
show the reaction diagram
-
methylates positions 2 and 7
-
-
?
S-adenosyl-L-methionine + uroporphyrinogen I
?
show the reaction diagram
-, Q51720
methylates positions 2 and 7
-
-
?
S-adenosyl-L-methionine + uroporphyrinogen III
?
show the reaction diagram
Lactobacillus reuteri, Lactobacillus reuteri CRL 1098
-
-
-
-
?
S-adenosyl-L-methionine + uroporphyrinogen III
S-adenosyl-L-homocysteine + precorrin-2
show the reaction diagram
P95417, -
-
-
-
ir
uroporphyrinogen III + S-adenosyl-L-methionine
precorrin-2 + S-adenosyl-L-homocysteine
show the reaction diagram
-
-
-
-
?
uroporphyrinogen III + S-adenosyl-L-methionine
precorrin-2 + S-adenosyl-L-homocysteine
show the reaction diagram
-
via precorrin-1
-
-
?
uroporphyrinogen III + S-adenosyl-L-methionine
precorrin-2 + S-adenosyl-L-homocysteine
show the reaction diagram
-
NirE binds S-adenosyl-L-methionine, although not with the same avidity as reported for other SAM-dependent uroporphyrinogen III methyltransferases involved in siroheme and cobalamin biosynthesis
-
-
?
uroporphyrinogen III + S-adenosyl-L-methionine
precorrin-2 + S-adenosyl-L-homocysteine
show the reaction diagram
-, Q6TA16
the enzyme binds S-adenosyl-L-methionine and catalyzes the production of III methyl uroporphyrinogen in vitro
-
-
?
uroporphyrinogen III + S-adenosyl-L-methionine
precorrin-2 + S-adenosyl-L-homocysteine
show the reaction diagram
Geobacillus stearothermophilus V
Q6TA16
the enzyme binds S-adenosyl-L-methionine and catalyzes the production of III methyl uroporphyrinogen in vitro
-
-
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
Pseudomonas denitrificans SC510
-
-, involved in biosynthesis of cobalamin
product is precorrin-2
?
additional information
?
-
-, Q96532
no formation of sirohydrochlorin in presence of NAD+ or NADP+
-
-
-
additional information
?
-
-
uroporphyrin III is not a substrate of the enzyme
-
-
-
additional information
?
-
-
uroporphyrinogen III synthase is fused to uroporphyrinogen III methyltransferase in Desulfovibrio vulgaris
-
-
-
additional information
?
-
-
at high enzyme concentrations NirE catalyzes an overmethylation of uroporphyrinogen III, resulting in the formation of trimethylpyrrocorphin
-
-
-
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
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
-
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
-
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
-
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
involved in biosynthesis of sirohydrochlorin
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
involved in biosynthesis of sirohydrochlorin
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-, Q51720
involved in biosynthesis of sirohydrochlorin
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
involved in biosynthesis of siroheme
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
involved in biosynthesis of siroheme
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-, Q96532
involved in biosynthesis of siroheme
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
involved in biosynthesis of cobalamin
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-
involved in biosynthesis of cobalamin
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
P21632
involved in biosynthesis of cobalamin
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-, Q51720
involved in biosynthesis of cobalamin
product is precorrin-2
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
-, Q96532
no synthesis of cobalamin
product is precorrin-2
?
uroporphyrinogen III + S-adenosyl-L-methionine
precorrin-2 + S-adenosyl-L-homocysteine
show the reaction diagram
-
-
-
-
?
uroporphyrinogen III + S-adenosyl-L-methionine
precorrin-2 + S-adenosyl-L-homocysteine
show the reaction diagram
-
via precorrin-1
-
-
?
2 S-adenosyl-L-methionine + uroporphyrinogen III
2 S-adenosyl-L-homocysteine + dihydrosirohydrochlorin + 2 H+
show the reaction diagram
Pseudomonas denitrificans SC510
-
involved in biosynthesis of cobalamin
product is precorrin-2
?
additional information
?
-
-
uroporphyrinogen III synthase is fused to uroporphyrinogen III methyltransferase in Desulfovibrio vulgaris
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
S-adenosyl-L-methionine
-
-
S-adenosyl-L-methionine
-
dependent on
S-adenosyl-L-methionine
-, Q6TA16
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Co2+
-
complete inhibition by 0.1 mM, mechanism unclear
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
S-adenosyl-L-homocysteine
-
competitive
S-adenosyl-L-homocysteine
-
product inhibition at concentrations above 0.002 mM
S-adenosyl-L-homocysteine
P95417, -
-
uroporphyrinogen III
-
substrate inhibition above 0.002 mM
uroporphyrinogen III
-
substrate inhibition at concentrations above 0.017 mM
additional information
-
no inhibition by cobalamine or intermediates of synthesis
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0042
-
heptacarboxyporphyrinogen III
-
-
-
0.0047
-
hexacarboxyporphyrinogen III
-
-
-
0.0034
-
pentacarboxyporphyrinogen III
-
-
-
0.0063
-
S-adenosyl-L-methionine
-
-
0.0004
-
uroporphyrinogen III
-
pH 8.0, 22°C, CobA/HemD
0.001
-
uroporphyrinogen III
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
38
-
uroporphyrinogen III
-
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.00032
-
S-adenosyl-L-homocysteine
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.000044
-
-
recombinant enzyme
0.003
-
-
purified recombinant CobA/HemD
0.0062
-
-
purified enzyme
0.0091
-
-, Q96532
purified enzyme
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
22
-
-
assay at room temperature
37
-
-
assay at
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
27000
-
-, Q51720
SDS-PAGE
27610
-
-
calculated from DNA sequence
29200
-
P21632
calculated from DNA sequence
30000
-
-
SDS-PAGE
39900
-
-, Q96532
calculated from DNA sequence
41000
-
-, Q96532
SDS-PAGE
49930
-
-
calculated from DNA-sequence
50000
-
-
gel filtration
52000
-
-
SDS-PAGE
52000
-
-
gel filtration
56000
-
-
gel filtration
60000
-
-
gel filtration
60000
-
-, Q6TA16
recombinant enzyme, native PAGE and gel filtration
66000
-
-
gel filtration
67000
-
-, Q96532
SDS-PAGE, uncleaved fusion protein
79000
-
-
dynamic light scattering assay
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 55000, about, recombinant CobA/HemD, SDS-PAGE
?
-
x * 51000, SDS-PAGE
?
Lactobacillus reuteri CRL 1098
-
x * 51000, SDS-PAGE
-
dimer
-
homodimer, 2 * 30000, SDS-PAGE, gel filtration
dimer
-
homodimer, 2 * 27000, SDS-PAGE, gel filtration
dimer
-
crystallization data, 2 * 26233, calculated
dimer
-
crystallization data
homodimer
-
2 * 34000, SDS-PAGE
homodimer
-
2 * 30000, SDS-PAGE
homodimer
-, Q6TA16
2 * 30000, recombinant enzyme, SDS-PAGE
homodimer
P95417, -
-
homodimer
Geobacillus stearothermophilus V
-
2 * 30000, recombinant enzyme, SDS-PAGE
-
monomer
-
SDS-PAGE, gel filtration
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
sitting drop vapor diffusion method, using Tris, pH 8.0, 24% (w/v) PEG 6000 for enzyme without substrate, and 0.2 M lithium sulfate, 0.1 M Tris, pH 8.5, 1.26 M ammonium sulfate for enzyme in complex with uroporphyrinogen III
P95417, -
enzyme topology diagramm
-
both apoform and with S-adenosyl-L-methionine
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
65
-
-
thermostable, 17% of activity remaining after 10 min incubation
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
complete loss of activity when treated with Triton X-100
-, Q96532
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
4°C, 10 mM Tris, pH 7.7, stable for several weeks
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
from recombinant E. coli
-, Q96532
recombinant CobA/HemD , HemD, and CobA from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
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from recombinant E. coli
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Ni2+-Sepharose column chromatography
P95417, -
recombinant His-tagged NirE from Escherichia coli by metal chelate chromatography and gel filtration to homogeneity
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from recombinant E. coli using His-tag
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recombinant protein
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by Ni–NTA column
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Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
in Escherichia coli CBK103, amino acids 202-457 responsible for EC 2.1.1.107 activity
-, Q96532
DNA and amino acid sequence determination and analysis, the uroporphyrinogen III synthase is naturally fused with the methyltransferase, bypassing the need for uroporphyrinogen III decarboxylase activity, overview. Expression of cobA/hemD and isolate hemD or cobA in Escherichia coli strain BL21(DE3)
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complementation experiments with cysteine autotroph strain
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cysG containing plasmid transformed into Escherichia coli CBK103; overexpression of cysG gene
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overexpression of cysG gene
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gene cobA, expression in Escherichia coli strain JM109(DE3), the recombinant enzyme renders the bacteria morre resistant to potassium tellurite and potassium tellurate
-, Q6TA16
in Escherichia coli JM83 and MV1184
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expressed in Escherichia coli BL21(DE3) cells
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overexpression of cobA gene in Escherichia coli TB1
-, Q51720
recombinant expression of C-terminally His-tagged NirE in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH10B
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50 fold increase of enzyme activity when overexpressed
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expression of 5 open reading frames, codon 1120-1959 (cobA) encodes EC 2.1.1.107 activity
P21632
EC 2.1.1.107 deficient mutants complemented with cysG from Salmonella typhimurium and Escherichia coli and cobA from Pseudomonas denitrificans, overexpression of yeast gene product in Escherichia coli
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expression in Escherichia coli
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Escherichia coli JM109 used for cloning all recombinant plasmids and expressing the pUC18-derived plasmid, Escherichia coli strain S13009 and BL21(DE3) used for expressing the pQE-derived plasmids and the pET-derived plasmid
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in Escherichia coli CBK103
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ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
D227A
-
full activity
D248A
-
no transmethylase activity
D303A
-
full activity
G224A
-
unable to bind S-adenosyl-L-methionine
K270I
-
full activity
R298L
-
unable to bind S-adenosyl-L-methionine
R309L
-
no transmethylase activity
G12A
-, Q6TA16
site-directed mutagenesis of cobA abolishes the tellurite resistance of the mesophilic, heterologous host Escherichia coli and the SUMT activity in vitro. Cells overexpressing SUMT G12A show 7fold less tolerance to K2TeO3 as compared to that exhibit by cells expressing the wild-type methyltransferase
G12A
Geobacillus stearothermophilus V
-
site-directed mutagenesis of cobA abolishes the tellurite resistance of the mesophilic, heterologous host Escherichia coli and the SUMT activity in vitro. Cells overexpressing SUMT G12A show 7fold less tolerance to K2TeO3 as compared to that exhibit by cells expressing the wild-type methyltransferase
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E114Q
P95417, -
the mutant shows strongly reduced activity compared to the wild type enzyme
G189K
P95417, -
the mutant shows increased activity and reduced S-adenosyl-L-methionine-binding ability compared to the wild type enzyme
G189N
P95417, -
the mutant shows increased activity and reduced S-adenosyl-L-methionine-binding ability compared to the wild type enzyme
H161F
P95417, -
the mutant shows reduced activity compared to the wild type enzyme
K102A
P95417, -
the mutant shows no activity and strongly reduced S-adenosyl-L-methionine-binding ability compared to the wild type enzyme
R111K
P95417, -
the mutant shows strongly reduced activity and reduced S-adenosyl-L-methionine-binding ability compared to the wild type enzyme
R149K
P95417, -
the mutant shows no activity compared to the wild type enzyme
R51K
P95417, -
the mutant shows reduced activity and reduced S-adenosyl-L-methionine-binding ability compared to the wild type enzyme
D47N
-
binds about a quarter the quantity of S-adenosyl-L-methionine compared with wild-type, reaction product is only poxidised precorrin-1
F106A
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considerably less soluble than wild-type, no binding of S-adenosyl-L-methionine, no enzymic activity
L49A
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binds about half the quantity of S-adenosyl-L-methionine compared with wild-type, reaction products are oxidised forms of both precorrin-1 and precorrin-2
M184A
-
slight enzymic activity
T130A
-
considerably less soluble than wild-type, no binding of S-adenosyl-L-methionine
additional information
-
separation of the individual enzyme activities, uroporphyrinogen III synthase and uroporphyrinogen III methyltransferase, by dissecting the relevant gene to allow the production of two distinct proteins
M186L
P95417, -
the mutant shows strongly reduced activity and reduced S-adenosyl-L-methionine-binding ability compared to the wild type enzyme
additional information
-
construction of a nirE transposon mutant, which is not complemented by native cobA encoding the SAM-dependent uroporphyrinogen III methyltransferase involved in cobalamin formation, overview
Y183A
-
considerably less soluble than wild-type, no binding of S-adenosyl-L-methionine, no enzyminc activity
additional information
-
deletion mutants, truncated one deleting the C-terminal extra 52 amino acids actively expressed in Escherichia coli, the mature SUMT fusion mutant or the mature SUMT deleting the N-terminal 36 amino acids including glycine-rich region involved directly in SAM binding expressed as an inclusion body in Escherichia coli
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
analysis
-
red fluorescent compounds are associated with the recombinant mature SUMT which are identified as sirohydrochlorin and trimethylpyrrocorphin by spectroscopic analysis, slightly alter the protein secondary structure