BRENDA - Enzyme Database show
show all sequences of 2.1.1.148

Functional evidence for active site location of tetrameric thymidylate synthase X at the interphase of three monomers

Leduc, D.; Graziani, S.; Lipowski, G.; Marchand, C.; Le Marechal, P.; Liebl, U.; Myllykallio, H.; Proc. Natl. Acad. Sci. USA 101, 7252-7257 (2004)

Data extracted from this reference:

Engineering
Amino acid exchange
Commentary
Organism
H48Q
inactive mutant enzyme
Helicobacter pylori
H48Q/S84A
inactive mutant enzyme
Helicobacter pylori
H48Q/S84C
inactive mutant enzyme
Helicobacter pylori
R74A
the KM-value for dUMP is 5.1fold higher than the wild-type value, the turnover-number is 4.6fold lower than the wild-type value
Helicobacter pylori
R74K
the KM-value for dUMP is nearly identical to wild-type value, the turnover-number is 6.6fold lower than the wild-type value
Helicobacter pylori
S84A
the KM-value for dUMP is 5.3fold higher than the wild-type value, the turnover-number is 2.1fold lower than the wild-type value. Mutation abolishes thymidylate synthase activity in vivo
Helicobacter pylori
S84A/S85A
inactive mutant enzyme
Helicobacter pylori
S84C
mutation abolishes thymidylate synthase activity in vivo
Helicobacter pylori
S84Y
the KM-value for dUMP is 1.9fold higher than the wild-type value, the turnover-number is 1.6fold lower than the wild-type value
Helicobacter pylori
Y87F
the KM-value for dUMP is 1.8fold lower than the wild-type value, the turnover-number is nearly identical to the wild-type value
Helicobacter pylori
Inhibitors
Inhibitors
Commentary
Organism
Structure
additional information
functional evidence for active site location of tetrameric thymidylate synthase X at the interphase of three monomers. The active-site configurations of ThyX proteins, present in many human pathogenic bacteria, and of human thymidylate synthase A (EC 2.1.1.45) are different.
Helicobacter pylori
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.00083
-
dUMP
mutant enzyme S84C
Helicobacter pylori
0.0012
-
dUMP
mutant enzyme R74K
Helicobacter pylori
0.0017
-
dUMP
mutant enzyme R74A
Helicobacter pylori
0.0037
-
dUMP
mutant enzyme S84A
Helicobacter pylori
0.0076
-
dUMP
wild-type enzyme
Helicobacter pylori
0.008
-
dUMP
mutant enzyme Y87F
Helicobacter pylori
0.012
-
dUMP
mutant enzyme S84Y
Helicobacter pylori
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Helicobacter pylori
-
-
-
Purification (Commentary)
Commentary
Organism
-
Helicobacter pylori
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
5,10-methylenetetrahydrofolate + dUMP + FADH2
-
660374
Helicobacter pylori
dTMP + tetrahydrofolate + FAD
-
-
-
?
Subunits
Subunits
Commentary
Organism
?
x * 27168.2, wild-type enzyme, MALDI-TOF
Helicobacter pylori
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.0038
-
dUMP
mutant enzyme Y87F
Helicobacter pylori
0.0072
-
dUMP
wild-type enzyme
Helicobacter pylori
0.0078
-
dUMP
mutant enzyme R74K
Helicobacter pylori
0.0097
-
dUMP
mutant enzyme S84Y
Helicobacter pylori
0.037
-
dUMP
mutant enzyme R74A
Helicobacter pylori
0.0384
-
dUMP
mutant enzyme S84A
Helicobacter pylori
0.14
-
dUMP
mutant enzyme S84C
Helicobacter pylori
Cofactor
Cofactor
Commentary
Organism
Structure
FAD
wild-type and mutant enzymes Y87F, S84Y, H48Q, S84A and S84C contain quantities of tightly bound FAD. Mutant enzymes R74A, R74K, H48Q/S84A and H48Q/S84C easily lose considerable amounts of bound FAD
Helicobacter pylori
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FAD
wild-type and mutant enzymes Y87F, S84Y, H48Q, S84A and S84C contain quantities of tightly bound FAD. Mutant enzymes R74A, R74K, H48Q/S84A and H48Q/S84C easily lose considerable amounts of bound FAD
Helicobacter pylori
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
H48Q
inactive mutant enzyme
Helicobacter pylori
H48Q/S84A
inactive mutant enzyme
Helicobacter pylori
H48Q/S84C
inactive mutant enzyme
Helicobacter pylori
R74A
the KM-value for dUMP is 5.1fold higher than the wild-type value, the turnover-number is 4.6fold lower than the wild-type value
Helicobacter pylori
R74K
the KM-value for dUMP is nearly identical to wild-type value, the turnover-number is 6.6fold lower than the wild-type value
Helicobacter pylori
S84A
the KM-value for dUMP is 5.3fold higher than the wild-type value, the turnover-number is 2.1fold lower than the wild-type value. Mutation abolishes thymidylate synthase activity in vivo
Helicobacter pylori
S84A/S85A
inactive mutant enzyme
Helicobacter pylori
S84C
mutation abolishes thymidylate synthase activity in vivo
Helicobacter pylori
S84Y
the KM-value for dUMP is 1.9fold higher than the wild-type value, the turnover-number is 1.6fold lower than the wild-type value
Helicobacter pylori
Y87F
the KM-value for dUMP is 1.8fold lower than the wild-type value, the turnover-number is nearly identical to the wild-type value
Helicobacter pylori
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
additional information
functional evidence for active site location of tetrameric thymidylate synthase X at the interphase of three monomers. The active-site configurations of ThyX proteins, present in many human pathogenic bacteria, and of human thymidylate synthase A (EC 2.1.1.45) are different.
Helicobacter pylori
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.00083
-
dUMP
mutant enzyme S84C
Helicobacter pylori
0.0012
-
dUMP
mutant enzyme R74K
Helicobacter pylori
0.0017
-
dUMP
mutant enzyme R74A
Helicobacter pylori
0.0037
-
dUMP
mutant enzyme S84A
Helicobacter pylori
0.0076
-
dUMP
wild-type enzyme
Helicobacter pylori
0.008
-
dUMP
mutant enzyme Y87F
Helicobacter pylori
0.012
-
dUMP
mutant enzyme S84Y
Helicobacter pylori
Purification (Commentary) (protein specific)
Commentary
Organism
-
Helicobacter pylori
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
5,10-methylenetetrahydrofolate + dUMP + FADH2
-
660374
Helicobacter pylori
dTMP + tetrahydrofolate + FAD
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
?
x * 27168.2, wild-type enzyme, MALDI-TOF
Helicobacter pylori
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.0038
-
dUMP
mutant enzyme Y87F
Helicobacter pylori
0.0072
-
dUMP
wild-type enzyme
Helicobacter pylori
0.0078
-
dUMP
mutant enzyme R74K
Helicobacter pylori
0.0097
-
dUMP
mutant enzyme S84Y
Helicobacter pylori
0.037
-
dUMP
mutant enzyme R74A
Helicobacter pylori
0.0384
-
dUMP
mutant enzyme S84A
Helicobacter pylori
0.14
-
dUMP
mutant enzyme S84C
Helicobacter pylori
Other publictions for EC 2.1.1.148
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)
733512
Alexandrova
Synthesis and evaluation of C- ...
Mycobacterium tuberculosis
Bioorg. Med. Chem.
23
7131-7137
2015
-
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1
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1
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733298
Becker
Substrate interaction dynamics ...
Paramecium bursaria Chlorella virus 1
Biochem. J.
459
37-45
2014
-
-
-
-
-
-
-
-
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4
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1
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1
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734029
Mishanina
Substrate activation in flavin ...
Thermotoga maritima, Thermotoga maritima ATCC 43589
J. Am. Chem. Soc.
136
10597-10600
2014
-
-
-
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5
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2
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-
-
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-
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-
-
-
734346
Mathews
Flavin-dependent thymidylate s ...
Thermotoga maritima, Thermotoga maritima ATCC 43589
J. Bioterror Biodef.
Suppl 12
004
2013
-
-
1
1
1
-
-
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-
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5
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1
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1
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-
719663
Mishanina
Trapping of an intermediate in ...
Thermotoga maritima, Thermotoga maritima ATCC 43589
J. Am. Chem. Soc.
134
4442-4448
2012
-
-
-
-
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5
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1
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1
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1
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2
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-
720985
Zhang
Crystal structure of a flavin- ...
Helicobacter pylori
Protein Pept. Lett.
19
1225-1230
2012
-
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-
1
-
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6
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1
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1
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1
1
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1
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-
-
-
-
-
-
-
-
-
-
-
720992
Wang
Crystal structure and enzymati ...
Helicobacter pylori, Helicobacter pylori SS1
Protein Sci.
20
1398-1410
2011
1
-
1
1
-
-
1
2
-
-
3
-
-
4
-
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1
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2
1
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1
1
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1
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1
1
1
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1
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3
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1
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2
1
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1
1
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-
718969
Kan
Biochemical characterization o ...
no activity in Corynebacterium glutamicum strain NCHU 87078
Biochim. Biophys. Acta
1804
1751-1759
2010
-
-
-
-
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1
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697906
Wang
Oxidase activity of a flavin-d ...
Thermotoga maritima
FEBS J.
276
2801-2810
2009
-
-
-
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1
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2
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1
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700395
Koehn
An unusual mechanism of thymid ...
Thermotoga maritima
Nature
458
919-923
2009
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1
2
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2
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1
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1
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-
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691285
Esra Onen
Design, synthesis and evaluati ...
Paramecium bursaria Chlorella virus 1
Bioorg. Med. Chem. Lett.
18
3628-3631
2008
-
-
-
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-
5
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1
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1
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1
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2
5
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2
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5
5
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1
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1
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694812
Hunter
Kinetics and ligand-binding pr ...
Mycobacterium tuberculosis
PLoS ONE
3
e2237
2008
-
1
1
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1
3
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1
2
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1
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1
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3
1
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1
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1
1
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1
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3
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2
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1
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3
1
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698579
Ulmer
Functional analysis of the Myc ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
J. Bacteriol.
190
2056-2064
2008
-
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-
-
1
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-
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-
10
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1
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-
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-
700990
Escartin
Flavin-dependent thymidylate s ...
Helicobacter pylori, Paramecium bursaria Chlorella virus 1
Proc. Natl. Acad. Sci. USA
105
9948-9952
2008
-
-
-
-
-
-
-
-
-
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4
-
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-
-
-
-
-
-
-
-
-
1
2
2
1
-
-
671555
Chernyshev
Thymidyl biosynthesis enzymes ...
Bacillus anthracis, Chlamydia trachomatis, Clostridium botulinum, Helicobacter pylori, Mycobacterium leprae, Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv, no activity in Homo sapiens, Paramecium bursaria Chlorella virus-1, Pyrococcus furiosus, Rickettsia sp., Thermotoga maritima, Treponema palladium
Appl. Microbiol. Biotechnol.
74
282-289
2007
-
1
-
3
-
-
2
-
-
-
-
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-
17
-
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3
1
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2
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1
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2
3
-
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2
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3
1
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691759
Chernyshev
The relationships between oxid ...
Thermotoga maritima
Chem. Commun. (Camb. )
27
2861-2863
2007
-
1
-
-
-
-
-
-
-
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-
1
-
1
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-
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1
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2
-
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1
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2
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1
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1
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-
-
-
-
-
-
692837
Leduc
Flavin-dependent thymidylate s ...
Rhodobacter capsulatus, Rhodobacter capsulatus MT1131
J. Bacteriol.
189
8537-8545
2007
-
-
-
-
-
-
-
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2
-
6
-
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4
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2
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2
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2
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4
-
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-
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-
678023
Kanai
Archaeal Pyrococcus furiosus t ...
Pyrococcus furiosus
Biochem. J.
393
373-379
2006
-
1
1
-
-
-
-
-
-
1
1
-
-
3
-
-
1
-
-
-
-
-
1
1
1
1
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1
1
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-
-
-
-
-
-
-
1
1
-
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-
-
1
-
-
-
-
1
1
1
1
-
-
-
-
-
-
-
-
-
-
-
-
679184
Mason
A lag-phase in the reduction o ...
Thermotoga maritima
Chem. Commun. (Camb. )
16
1781-1783
2006
-
-
-
-
-
-
-
1
-
-
-
-
-
1
-
-
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-
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1
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1
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1
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1
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-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
680675
Graziani
Catalytic mechanism and struct ...
Paramecium bursaria Chlorella virus-1
J. Biol. Chem.
281
24048-24057
2006
-
-
1
1
9
-
2
-
-
-
-
-
-
4
-
-
1
-
-
-
-
1
1
1
-
-
-
-
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-
2
-
-
2
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1
2
1
9
-
2
2
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-
-
-
-
-
-
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1
-
-
-
1
1
1
-
-
-
-
-
-
-
-
-
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-
-
681384
Sampathkumar
NADP+ expels both the co-facto ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv
J. Mol. Biol.
360
1-6
2006
-
1
1
1
1
-
-
-
-
-
-
-
-
7
-
-
1
-
-
-
-
-
1
1
-
-
-
-
-
-
-
2
-
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-
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1
1
2
1
1
-
-
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1
-
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-
-
1
1
-
-
-
-
-
-
-
-
-
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-
-
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-
658992
Gattis
Direct observation of the part ...
Campylobacter jejuni
J. Am. Chem. Soc.
127
832-833
2005
-
-
-
-
-
-
-
-
-
-
-
-
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1
-
-
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-
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1
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2
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2
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-
1
-
-
-
-
-
-
-
-
-
-
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-
-
-
-
681375
Sampathkumar
Structure of the Mycobacterium ...
Mycobacterium tuberculosis
J. Mol. Biol.
352
1091-1104
2005
-
1
1
1
11
-
-
-
-
-
-
-
-
7
-
-
1
-
-
-
-
1
1
1
-
-
-
-
-
-
-
1
-
-
-
-
1
1
1
1
11
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
1
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
657899
Leduc
Two distinct pathways for thym ...
Aeropyrum pernix, Aquifex aeolicus, no activity in Archaeoglobus fulgidus, no activity in Methanocaldococcus jannaschii, no activity in Methanopyrus kandleri, no activity in Methanopyrus kandleri AV19, no activity in Methanothermobacter thermoautotrophicus, Pyrobaculum aerophilum, Pyrococcus abyssi, Pyrococcus furiosus, Pyrococcus horikoshii, Saccharolobus solfataricus, Sulfurisphaera tokodaii, Thermoplasma acidophilum, Thermoplasma volcanium, Thermotoga maritima
Biochem. Soc. Trans.
32
231-235
2004
-
-
-
-
-
-
-
-
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-
-
17
-
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-
-
-
-
-
-
22
-
-
-
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-
-
22
-
-
-
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-
-
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-
-
-
-
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-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
658015
Agrawal
Mechanistic studies of a flavi ...
Thermotoga maritima, Thermotoga maritima DSM 3109
Biochemistry
43
10295-10301
2004
-
1
-
-
-
-
1
-
-
-
-
1
-
6
-
-
-
-
-
-
-
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1
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-
-
-
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-
2
-
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-
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1
-
2
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1
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-
1
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-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
659052
Liu
Bacterial thymidylate synthase ...
Trichodesmium erythraeum
J. Bacteriol.
186
6316-6319
2004
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
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-
-
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-
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-
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1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
659429
Graziani
Functional analysis of FAD-dep ...
Paramecium bursaria Chlorella virus-1
J. Biol. Chem.
279
54340-54347
2004
-
-
1
-
1
-
5
2
-
-
1
-
-
5
-
-
1
-
-
-
-
-
2
1
-
-
-
-
-
-
-
3
-
-
-
-
-
1
3
-
1
-
-
5
-
2
-
-
1
-
-
-
-
1
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
660374
Leduc
Functional evidence for active ...
Helicobacter pylori
Proc. Natl. Acad. Sci. USA
101
7252-7257
2004
-
-
-
-
10
-
1
7
-
-
-
-
-
4
-
-
1
-
-
-
-
-
1
1
-
-
-
7
-
-
-
1
-
-
-
-
-
-
1
-
10
-
-
1
-
7
-
-
-
-
-
-
-
1
-
-
-
-
1
1
-
-
-
7
-
-
-
-
-
-
-
-
-
-
637234
Montfort
Complementing thymidylate synt ...
Dictyostelium discoideum, Thermotoga maritima
Structure
11
607-608
2003
4
-
-
-
-
-
-
-
-
-
-
2
-
2
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
2
-
-
-
4
-
-
2
-
-
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-
-
-
-
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-
2
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
637235
Mathews
Functional analysis of substra ...
Bacillus anthracis, Borreliella burgdorferi, Campylobacter jejuni, Chlamydia pneumoniae, Chlamydia psittaci, Chlamydia trachomatis, Clostridioides difficile, Clostridium botulinum, Clostridium perfringens, Corynebacterium diphtheriae, Dictyostelium discoideum, Ehrlichia chaffeensis, Helicobacter pylori, Mycobacterium avium, Mycobacterium leprae, Mycobacterium tuberculosis, Mycobacterium tuberculosis variant bovis, Rhodococcus sp., Rickettsia conorii, Rickettsia prowazekii, Thermotoga maritima, Thermotoga maritima DSM 3109, Treponema denticola, Treponema pallidum
Structure
11
677-690
2003
6
21
1
1
-
-
2
-
-
-
-
24
-
26
-
-
1
-
-
-
-
-
71
2
-
-
-
-
-
-
-
46
-
-
-
6
22
1
46
1
-
-
-
2
-
-
-
-
-
24
-
-
-
1
-
-
-
-
71
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
637232
Myllykallio
An alternative flavin-dependen ...
Borreliella burgdorferi, Campylobacter jejuni, Chlamydia sp., Dictyostelium discoideum, Helicobacter pylori, Mycobacterium tuberculosis, Pyrococcus abyssi, Pyrococcus horikoshii, Rickettsia prowazekii, Saccharolobus solfataricus, Treponema pallidum
Science
297
105-107
2002
3
4
2
-
1
-
-
-
-
-
3
11
-
11
-
-
1
-
-
-
-
-
33
1
-
-
-
-
-
-
-
22
-
-
-
3
4
2
22
-
1
-
-
-
-
-
-
-
3
11
-
-
-
1
-
-
-
-
33
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
637233
Giladi
Genetic evidence for a novel t ...
Campylobacter jejuni, Halobacterium salinarum
FEMS Microbiol. Lett.
216
105-109
2002
-
-
2
-
-
-
1
-
-
-
1
2
-
2
-
-
1
-
-
-
-
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4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
1
-
-
-
-
1
2
-
-
-
1
-
-
-
-
4
-
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-