BRENDA - Enzyme Database show
show all sequences of 1.16.1.8

Proximal FAD histidine residue influences interflavin electron transfer in cytochrome P450 reductase and methionine synthase reductase

Meints, C.E.; Parke, S.M.; Wolthers, K.R.; Arch. Biochem. Biophys. 547, 18-26 (2014)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain Rosetta2(DE3)pLysS
Homo sapiens
Engineering
Amino acid exchange
Commentary
Organism
A312H
site-directed mutangenesis, mutation of the catalytic residue leads to the kinetic coupling of hydride and interflavin electron transfer, and eliminates the formation of the FAD hydroquinone intermediate, substitution of Ala312 for His in MSR weakens NADP(H) binding as the Km for NADPH and Ki for NADP+ increases 6 and 1.7fold, respectively. NADPH reduction of A312H resembles that of native cytochrome P450 reductase, in that it occurs in two discrete kinetic phases, without the transient formation of the E-FADH2-FMN intermediate
Homo sapiens
A312Q
site-directed mutangenesis, the catalytic site mutant shows a 2.5fold increased Km and a slightly decreased Ki for the coenzyme FAD
Homo sapiens
Inhibitors
Inhibitors
Commentary
Organism
Structure
NADP+
-
Homo sapiens
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
stopped-flow and steady-state kinetic analysis
Homo sapiens
0.0024
-
NADPH
pH 7.5, 25C, recombinant wild-type enzyme
Homo sapiens
0.0062
-
NADPH
pH 7.5, 25C, recombinant mutant A312Q
Homo sapiens
0.015
-
NADPH
pH 7.5, 25C, recombinant mutant A312H
Homo sapiens
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2 [methionine synthase]-cob(II)alamin + NADPH + H+ + 2 S-adenosyl-L-methionine
Homo sapiens
-
2 [methionine synthase]-methylcob(I)alamin + 2 S-adenosylhomocysteine + NADP+
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Homo sapiens
Q9UBK8
-
-
Reaction
Reaction
Commentary
Organism
2 [methionine synthase]-methylcob(I)alamin + 2 S-adenosylhomocysteine + NADP+ = 2 [methionine synthase]-cob(II)alamin + NADPH + H+ + 2 S-adenosyl-L-methionine
hydride transfer and interflavin electron transfer are two catalytic steps represented by two distinct kinetic phases leading to transient formation of the FAD hydroquinone
Homo sapiens
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2 [methionine synthase]-cob(II)alamin + NADPH + H+ + 2 S-adenosyl-L-methionine
-
741764
Homo sapiens
2 [methionine synthase]-methylcob(I)alamin + 2 S-adenosylhomocysteine + NADP+
-
-
-
?
Temperature Optimum [C]
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
25
-
assay at
Homo sapiens
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.6
-
NADPH
pH 7.5, 25C, recombinant mutant A312H
Homo sapiens
7.2
-
NADPH
pH 7.5, 25C, recombinant wild-type enzyme
Homo sapiens
7.8
-
NADPH
pH 7.5, 25C, recombinant mutant A312Q
Homo sapiens
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Homo sapiens
Cofactor
Cofactor
Commentary
Organism
Structure
FAD
the proximal FAD histidine residue accelerates proton-coupled electron transfer from FADH2 to the higher potential FMN
Homo sapiens
FMN
the proximal FAD histidine residue accelerates proton-coupled electron transfer from FADH2 to the higher potential FMN
Homo sapiens
additional information
mechanism of NADPH reduction of a diflavin enzyme, overview
Homo sapiens
NADPH
-
Homo sapiens
Ki Value [mM]
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.0291
-
NADP+
pH 7.5, 25C, recombinant mutant A312Q
Homo sapiens
0.0369
-
NADP+
pH 7.5, 25C, recombinant wild-type enzyme
Homo sapiens
0.0729
-
NADP+
pH 7.5, 25C, recombinant mutant A312H
Homo sapiens
Cloned(Commentary) (protein specific)
Commentary
Organism
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain Rosetta2(DE3)pLysS
Homo sapiens
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FAD
the proximal FAD histidine residue accelerates proton-coupled electron transfer from FADH2 to the higher potential FMN
Homo sapiens
FMN
the proximal FAD histidine residue accelerates proton-coupled electron transfer from FADH2 to the higher potential FMN
Homo sapiens
additional information
mechanism of NADPH reduction of a diflavin enzyme, overview
Homo sapiens
NADPH
-
Homo sapiens
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
A312H
site-directed mutangenesis, mutation of the catalytic residue leads to the kinetic coupling of hydride and interflavin electron transfer, and eliminates the formation of the FAD hydroquinone intermediate, substitution of Ala312 for His in MSR weakens NADP(H) binding as the Km for NADPH and Ki for NADP+ increases 6 and 1.7fold, respectively. NADPH reduction of A312H resembles that of native cytochrome P450 reductase, in that it occurs in two discrete kinetic phases, without the transient formation of the E-FADH2-FMN intermediate
Homo sapiens
A312Q
site-directed mutangenesis, the catalytic site mutant shows a 2.5fold increased Km and a slightly decreased Ki for the coenzyme FAD
Homo sapiens
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
NADP+
-
Homo sapiens
Ki Value [mM] (protein specific)
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
0.0291
-
NADP+
pH 7.5, 25C, recombinant mutant A312Q
Homo sapiens
0.0369
-
NADP+
pH 7.5, 25C, recombinant wild-type enzyme
Homo sapiens
0.0729
-
NADP+
pH 7.5, 25C, recombinant mutant A312H
Homo sapiens
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
additional information
-
additional information
stopped-flow and steady-state kinetic analysis
Homo sapiens
0.0024
-
NADPH
pH 7.5, 25C, recombinant wild-type enzyme
Homo sapiens
0.0062
-
NADPH
pH 7.5, 25C, recombinant mutant A312Q
Homo sapiens
0.015
-
NADPH
pH 7.5, 25C, recombinant mutant A312H
Homo sapiens
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2 [methionine synthase]-cob(II)alamin + NADPH + H+ + 2 S-adenosyl-L-methionine
Homo sapiens
-
2 [methionine synthase]-methylcob(I)alamin + 2 S-adenosylhomocysteine + NADP+
-
-
?
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2 [methionine synthase]-cob(II)alamin + NADPH + H+ + 2 S-adenosyl-L-methionine
-
741764
Homo sapiens
2 [methionine synthase]-methylcob(I)alamin + 2 S-adenosylhomocysteine + NADP+
-
-
-
?
Temperature Optimum [C] (protein specific)
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
25
-
assay at
Homo sapiens
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
3.6
-
NADPH
pH 7.5, 25C, recombinant mutant A312H
Homo sapiens
7.2
-
NADPH
pH 7.5, 25C, recombinant wild-type enzyme
Homo sapiens
7.8
-
NADPH
pH 7.5, 25C, recombinant mutant A312Q
Homo sapiens
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.5
-
assay at
Homo sapiens
Other publictions for EC 1.16.1.8
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
745764
Cheng
Functional variant in methion ...
Homo sapiens
Mol. Cell. Biochem.
407
51-56
2015
-
-
1
-
-
-
-
-
-
-
-
-
-
1
-
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1
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-
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-
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-
-
-
-
-
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1
1
-
-
-
741764
Meints
Proximal FAD histidine residu ...
Homo sapiens
Arch. Biochem. Biophys.
547
18-26
2014
-
-
1
-
2
-
1
4
-
-
-
1
-
2
-
-
-
1
-
-
-
-
1
-
1
-
-
3
1
-
-
4
3
-
-
-
-
1
4
-
2
-
-
1
3
4
-
-
-
1
-
-
-
-
-
-
-
-
1
-
1
-
-
3
1
-
-
-
-
-
-
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-
744874
Haque
Distinct conformational behav ...
Homo sapiens
FEBS J.
281
5325-5340
2014
-
-
-
-
-
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1
-
-
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1
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1
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2
1
1
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4
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4
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1
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1
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2
1
1
-
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-
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-
-
745034
Garcia-Minguillan
Riboflavin status modifies th ...
Homo sapiens
Genes Nutr.
9
435
2014
-
-
1
-
1
-
-
-
-
-
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1
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1
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1
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4
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1
4
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1
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1
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-
1
-
-
-
-
-
-
-
-
-
-
3
3
-
-
-
727476
Meints
Aromatic substitution of the F ...
Homo sapiens
FEBS J.
280
1460-1474
2013
-
-
-
-
2
-
2
1
-
-
-
1
-
1
-
-
-
1
-
-
-
-
3
-
1
-
-
-
1
-
-
3
6
-
-
-
-
-
3
-
2
-
-
2
6
1
-
-
-
1
-
-
-
-
-
-
-
-
3
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
716185
Han
Methionine synthase reductase ...
Homo sapiens
Mol. Biol. Rep.
39
805-816
2012
-
1
1
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
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-
1
1
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1
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-
-
-
-
-
-
-
-
-
-
-
-
-
2
2
-
-
-
727334
Zhao
A functional variant in MTRR i ...
Homo sapiens
Circulation
125
482-490
2012
-
-
1
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
1
-
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1
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1
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1
2
2
1
-
-
714738
Rigby
ELDOR spectroscopy reveals tha ...
Homo sapiens
ChemBioChem
12
863-867
2011
-
-
-
-
-
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-
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-
-
-
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2
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1
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1
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1
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1
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-
726959
Meints
Tryptophan 697 modulates hydri ...
Homo sapiens
Biochemistry
50
11131-11142
2011
-
-
-
-
4
-
2
1
-
-
-
1
-
2
-
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-
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-
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3
-
1
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1
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3
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3
-
4
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2
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1
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1
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3
-
1
-
-
-
1
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-
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-
1
1
-
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-
676037
Elmore
Metabolic derangement of methi ...
Mus musculus
Mol. Genet. Metab.
91
85-97
2007
-
-
-
-
1
-
-
-
-
-
-
-
-
4
-
-
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1
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685093
Wolthers
Mechanism of coenzyme binding ...
Homo sapiens
Biochemistry
46
11833-11844
2007
-
-
-
1
-
-
2
2
-
-
1
2
-
2
-
-
-
-
-
-
-
-
4
2
1
-
-
1
1
-
-
5
3
-
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-
-
5
1
-
-
-
2
3
2
-
-
1
2
-
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-
4
2
1
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1
1
-
-
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-
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-
685164
Wolthers
Protein interactions in the hu ...
Homo sapiens
Biochemistry
46
6696-6709
2007
-
-
1
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1
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1
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3
-
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1
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2
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1
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1
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4
-
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1
4
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1
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1
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1
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2
-
1
-
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1
-
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-
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-
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-
673482
Kim
Methionine synthase reductase ...
Homo sapiens
Exp. Mol. Med.
38
519-524
2006
-
1
-
-
2
-
-
-
-
-
-
-
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2
-
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1
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1
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1
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676875
Yamada
Human methionine synthase redu ...
Homo sapiens
Proc. Natl. Acad. Sci. USA
103
9476-9481
2006
-
-
-
-
-
-
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3
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1
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1
1
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1
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1
1
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658055
Olteanu
Kinetic and thermodynamic char ...
Homo sapiens
Biochemistry
43
1988-1997
2004
-
1
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2
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1
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657920
Olteanu
Differences in the efficiency ...
Homo sapiens
Biochemistry
41
13378-13385
2002
-
-
-
-
-
-
-
10
-
-
-
1
-
2
-
-
-
-
-
-
-
-
6
-
-
-
-
5
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
10
-
-
-
1
-
-
-
-
-
-
-
-
6
-
-
-
-
5
-
-
-
-
-
-
-
-
-
-
659159
Olteanu
Human methionine synthase redu ...
Homo sapiens
J. Biol. Chem.
276
35558-35563
2001
-
-
1
-
-
-
-
1
1
-
2
-
-
4
-
-
1
-
-
-
1
-
2
1
-
-
-
-
-
-
-
4
-
-
-
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-
1
4
-
-
-
-
-
-
1
1
-
2
-
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-
-
1
-
-
1
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
659955
Wilson
A common variant in methionine ...
Homo sapiens
Mol. Genet. Metab.
67
317-323
1999
-
1
-
-
-
-
-
-
-
-
-
-
-
2
-
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484889
Leclerc
Cloning and mapping of a cDNA ...
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3059-3064
1998
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484890
Jarrett
The mechanism of adenosylmethi ...
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37
12649-12658
1998
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