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Information on EC 2.4.2.18 - anthranilate phosphoribosyltransferase and Organism(s) Saccharolobus solfataricus and UniProt Accession P50384
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2.4.2.18 anthranilate phosphoribosyltransferaseIUBMB Comments
In some organisms, this enzyme is part of a multifunctional protein together with one or more other components of the system for biosynthesis of tryptophan [EC 4.1.1.48 (indole-3-glycerol-phosphate synthase), EC 4.1.3.27 (anthranilate synthase), EC 4.2.1.20 (tryptophan synthase) and EC 5.3.1.24 (phosphoribosylanthranilate isomerase)].
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Word Map
- 2.4.2.18
-
phosphoribosyltransferases
- chorismate
-
sulfolobus
- solfataricus
- phosphorylases
-
indole-3-glycerol
-
glutamine-dependent
- hafniae
-
4.1.3.27
The taxonomic range for the selected organisms is: Saccharolobus solfataricus
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
Synonyms
anthranilate phosphoribosyltransferase, anprt, anthranilate phosphoribosyl transferase, phosphoribosylanthranilate transferase, ssanprt, sanprt, sstrpd, anthranilate prt, trp d, anthranilate 5-phosphoribosylpyrophosphate phosphoribosyltransferase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
anthranilate 5-phosphoribosylpyrophosphate phosphoribosyltransferase
-
-
-
-
anthranilate phosphoribosylpyrophosphate phosphoribosyltransferase
-
-
-
-
anthranilate-5-phosphoribosylphosphate phosphoribosyltransferase
-
-
-
-
anthranilate-PP-ribose-P phosphoribosyltransferase
-
-
-
-
phosphoribosyl-anthranilate pyrophosphorylase
-
-
-
-
phosphoribosylanthranilate pyrophosphorylase
-
-
-
-
phosphoribosylanthranilate transferase
-
-
-
-
phosphoribosyltransferase, anthranilate
-
-
-
-
PR transferase
-
-
-
-
PRT
-
-
-
-
TrpD
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pentosyl group transfer
-
-
-
-
PATHWAY SOURCE
PATHWAYS
BRENDA
-
-
SYSTEMATIC NAME
IUBMB Comments
N-(5-phospho-D-ribosyl)-anthranilate:diphosphate phospho-alpha-D-ribosyltransferase
In some organisms, this enzyme is part of a multifunctional protein together with one or more other components of the system for biosynthesis of tryptophan [EC 4.1.1.48 (indole-3-glycerol-phosphate synthase), EC 4.1.3.27 (anthranilate synthase), EC 4.2.1.20 (tryptophan synthase) and EC 5.3.1.24 (phosphoribosylanthranilate isomerase)].
CAS REGISTRY NUMBER
COMMENTARY
9059-35-2
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
anthranilate + 5-phospho-alpha-D-ribose 1-diphosphate
N-(5-phospho-D-ribosyl)-anthranilate + diphosphate
anthranilate + 5-phospho-alpha-D-ribose 1-diphosphate
N-(5-phospho-D-ribosyl)-anthranilate + diphosphate
anthranilate + 5-phospho-alpha-D-ribose 1-diphosphate
N-(5-phospho-D-ribosyl)-anthranilate + diphosphate
-
-
-
?
anthranilate + 5-phospho-alpha-D-ribose 1-diphosphate
N-(5-phospho-D-ribosyl)-anthranilate + diphosphate
-
-
-
-
?
anthranilate + 5-phospho-alpha-D-ribose 1-diphosphate
N-(5-phospho-D-ribosyl)-anthranilate + diphosphate
-
-
-
?
anthranilate + 5-phospho-alpha-D-ribose 1-diphosphate
N-(5-phospho-D-ribosyl)-anthranilate + diphosphate
-
-
-
-
?
anthranilate + 5-phospho-alpha-D-ribose 1-diphosphate
N-(5-phospho-D-ribosyl)-anthranilate + diphosphate
-
-
-
?
anthranilate + 5-phospho-alpha-D-ribose 1-diphosphate
N-(5-phospho-D-ribosyl)-anthranilate + diphosphate
tryptophan biosynthesis, class III phosphoribosyltransfer
-
-
ir
anthranilate + 5-phospho-alpha-D-ribose 1-diphosphate
N-(5-phospho-D-ribosyl)-anthranilate + diphosphate
37°C, pH 7.2, 0.05 mM Mg2+
-
-
ir
anthranilate + 5-phospho-alpha-D-ribose 1-diphosphate
N-(5-phospho-D-ribosyl)-anthranilate + diphosphate
catalyzes the third step in tryptophan biosynthesis
-
-
?
anthranilate + 5-phospho-alpha-D-ribose 1-diphosphate
N-(5-phospho-D-ribosyl)-anthranilate + diphosphate
-
-
-
?
anthranilate + 5-phospho-alpha-D-ribose 1-diphosphate
N-(5-phospho-D-ribosyl)-anthranilate + diphosphate
enzyme of tryptophan biosynthesis
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
anthranilate + 5-phospho-alpha-D-ribose 1-diphosphate
N-(5-phospho-D-ribosyl)-anthranilate + diphosphate
anthranilate + 5-phospho-alpha-D-ribose 1-diphosphate
N-(5-phospho-D-ribosyl)-anthranilate + diphosphate
enzyme of tryptophan biosynthesis
-
-
?
anthranilate + 5-phospho-alpha-D-ribose 1-diphosphate
N-(5-phospho-D-ribosyl)-anthranilate + diphosphate
-
-
-
?
anthranilate + 5-phospho-alpha-D-ribose 1-diphosphate
N-(5-phospho-D-ribosyl)-anthranilate + diphosphate
tryptophan biosynthesis, class III phosphoribosyltransfer
-
-
ir
anthranilate + 5-phospho-alpha-D-ribose 1-diphosphate
N-(5-phospho-D-ribosyl)-anthranilate + diphosphate
catalyzes the third step in tryptophan biosynthesis
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Mg2+
marked inhibition at MgCl2 concentrations higher than 0.1 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.000005
anthranilate
at 87°C, Km decreases at lower temperatures
0.00007
5-phospho-alpha-D-ribose 1-diphosphate
60°C, pH 7.5, 2 mM Mg2+, mutant enzyme D223N
0.00016
5-phospho-alpha-D-ribose 1-diphosphate
60°C, pH 7.5, 0.05 mM Mg2+, wild-type enzyme
0.00017
5-phospho-alpha-D-ribose 1-diphosphate
60°C, pH 7.5, 0.05 mM Mg2+, mutant enzyme R164A
0.00026
5-phospho-alpha-D-ribose 1-diphosphate
60°C, pH 7.5, 2 mM Mg2+, mutant enzyme K106Q
0.0003
5-phospho-alpha-D-ribose 1-diphosphate
60°C, pH 7.5, 0.05 mM Mg2+, mutant enzyme H107A
0.0026
5-phospho-alpha-D-ribose 1-diphosphate
60°C, pH 7.5, 2 mM Mg2+, mutant enzyme E224Q
0.0043
5-phospho-alpha-D-ribose 1-diphosphate
60°C, pH 7.5, 2 mM Mg2+, wild-type enzyme
0.013
5-phospho-alpha-D-ribose 1-diphosphate
mutant D83G/F149S, in the presence of 0.05 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
0.023
5-phospho-alpha-D-ribose 1-diphosphate
mutant D83G/F149S, in the presence of 2 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
0.032
5-phospho-alpha-D-ribose 1-diphosphate
at pH 7.5 and 25°C
0.033
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/D83G/N109S/F149S, pH 7.2, 37°C
0.037
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/D83G/F149S/F193S, pH 7.2, 37°C
0.038
5-phospho-alpha-D-ribose 1-diphosphate
mutant D83G, in the presence of 2 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
0.045
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/D83G/F149S, pH 7.2, 37°C
0.047
5-phospho-alpha-D-ribose 1-diphosphate
wild-type, in the presence of 0.05 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
0.047
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/T77I/D83G/F149S/N109S/I169T/F193S/L320M, pH 7.2, 37°C
0.054
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/D83G/F149S/L320M, pH 7.2, 37°C
0.055
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/T77I/D83G/F149S, pH 7.2, 37°C
0.055
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/T77I/D83G/F149S/I169T/F193S/L320M, pH 7.2, 37°C
0.062
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/D83G/F149S/I169T, pH 7.2, 37°C
0.15
5-phospho-alpha-D-ribose 1-diphosphate
I36E/M47D double mutant, pH 7.2, 37°C, 0.01 microM subunit concentration
0.151
5-phospho-alpha-D-ribose 1-diphosphate
mutant F149S, in the presence of 0.05 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
0.18
5-phospho-alpha-D-ribose 1-diphosphate
wild-type, pH 7.2, 37°C, 0.01 microM subunit concentration
1.54
5-phospho-alpha-D-ribose 1-diphosphate
wild-type, in the presence of 2 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
0.000018
anthranilate
wild-type, in the presence of 0.05 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
0.00002
anthranilate
wild-type, in the presence of 2 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
0.00004
anthranilate
60°C, pH 7.5, 0.05 mM Mg2+, wild-type enzyme
0.000058
anthranilate
I36E/M47D double mutant, pH 7.2, 37°C, 0.01 microM subunit concentration
0.00007
anthranilate
60°C, pH 7.5, 2 mM Mg2+, wild-type enzyme
0.000085
anthranilate
wild-type, pH 7.2, 37°C, 0.01 microM subunit concentration
0.00012
anthranilate
mutant D83G, in the presence of 2 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
0.00016
anthranilate
60°C, pH 7.5, 2 mM Mg2+,mutant enzyme D223N
0.00053
anthranilate
60°C, pH 7.5, 0.05 mM Mg2+, mutant enzyme H107A
0.00075
anthranilate
60°C, pH 7.5, 2 mM Mg2+, mutant enzyme K106Q
0.00091
anthranilate
60°C, pH 7.5, 2 mM Mg2+,mutant enzyme E224Q
0.0012
anthranilate
mutant F149S, in the presence of 0.05 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
0.0031
anthranilate
mutant D83G/F149S, in the presence of 2 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
0.0036
anthranilate
mutant D83G/F149S, in the presence of 0.05 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
0.007
anthranilate
mutant enzyme I36E/M47D/D83G/F149S/I169T, pH 7.2, 37°C
0.008
anthranilate
mutant enzyme I36E/M47D/D83G/F149S/L320M, pH 7.2, 37°C
0.011
anthranilate
mutant enzyme I36E/M47D/D83G/F149S, pH 7.2, 37°C
0.0125
anthranilate
60°C, pH 7.5, 0.05 mM Mg2+, mutant enzyme H107A/P178A
0.029
anthranilate
mutant enzyme I36E/M47D/D83G/F149S/F193S, pH 7.2, 37°C
0.037
anthranilate
mutant enzyme I36E/M47D/T77I/D83G/F149S, pH 7.2, 37°C
0.051
anthranilate
mutant enzyme I36E/M47D/T77I/D83G/F149S/N109S/I169T/F193S/L320M, pH 7.2, 37°C
0.06
anthranilate
mutant enzyme I36E/M47D/D83G/N109S/F149S, pH 7.2, 37°C
0.082
anthranilate
mutant enzyme I36E/M47D/T77I/D83G/F149S/I169T/F193S/L320M, pH 7.2, 37°C
0.149
anthranilate
60°C, pH 7.5, 0.05 mM Mg2+,mutant enzyme R164A/H154A
0.297
anthranilate
60°C, pH 7.5, 0.05 mM Mg2+, mutant enzyme R164A
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.046
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/T77I/D83G/F149S/I169T/F193S/L320M, pH 7.2, 37°C
0.18
5-phospho-alpha-D-ribose 1-diphosphate
at pH 7.5 and 25°C
0.48
5-phospho-alpha-D-ribose 1-diphosphate
60°C, pH 7.5, 2 mM Mg2+, mutant enzyme K106Q
0.62
5-phospho-alpha-D-ribose 1-diphosphate
60°C, pH 7.5, 0.05 mM Mg2+, mutant enzyme R164A
0.73
5-phospho-alpha-D-ribose 1-diphosphate
60°C, pH 7.5, 0.05 mM Mg2+, mutant enzyme R164A/H154A
0.78
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/D83G/N109S/F149S, pH 7.2, 37°C
1.1
5-phospho-alpha-D-ribose 1-diphosphate
60°C, pH 7.5, 2 mM Mg2+, wild-type enzyme
1.2
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/T77I/D83G/F149S/N109S/I169T/F193S/L320M, pH 7.2, 37°C
1.3
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/T77I/D83G/F149S, pH 7.2, 37°C
2
5-phospho-alpha-D-ribose 1-diphosphate
60°C, pH 7.5, 0.05 mM Mg2+, mutant enzyme H107A/P178A
2.7
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/D83G/F149S/F193S, pH 7.2, 37°C
3.4
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/D83G/F149S, pH 7.2, 37°C
3.5
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/D83G/F149S/I169T, pH 7.2, 37°C
3.9
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/D83G/F149S/L320M, pH 7.2, 37°C
4.2
5-phospho-alpha-D-ribose 1-diphosphate
60°C, pH 7.5, 0.05 mM Mg2+, wild-type enzyme
5.1
5-phospho-alpha-D-ribose 1-diphosphate
60°C, pH 7.5, 0.05 mM Mg2+, mutant enzyme H107A
5.6
5-phospho-alpha-D-ribose 1-diphosphate
60°C, pH 7.5, 2 mM Mg2+, mutant enzyme E224Q
6
5-phospho-alpha-D-ribose 1-diphosphate
60°C, pH 7.5, 2 mM Mg2+, mutant enzyme D223N
0.046
anthranilate
mutant enzyme I36E/M47D/T77I/D83G/F149S/I169T/F193S/L320M, pH 7.2, 37°C
0.24
anthranilate
wild-type, in the presence of 2 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
0.28
anthranilate
+/-0.06, I36E/M47D double mutant, pH 7.2, 37°C, 0.01 microM subunit concentration
0.33
anthranilate
wild-type, in the presence of 0.05 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
0.41
anthranilate
+/-0.04, wild-type, pH 7.2, 37°C, 0.01 microM subunit concentration
0.48
anthranilate
60°C, pH 7.5, 2 mM Mg2+, mutant enzyme K106Q
0.62
anthranilate
60°C, pH 7.5, 0.05 mM Mg2+, mutant enzyme R164A
0.73
anthranilate
60°C, pH 7.5, 0.05 mM Mg2+,mutant enzyme R164A/H154A
0.78
anthranilate
mutant enzyme I36E/M47D/D83G/N109S/F149S, pH 7.2, 37°C
1.2
anthranilate
mutant F149S, in the presence of 0.05 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
1.2
anthranilate
mutant enzyme I36E/M47D/T77I/D83G/F149S/N109S/I169T/F193S/L320M, pH 7.2, 37°C
1.3
anthranilate
mutant enzyme I36E/M47D/T77I/D83G/F149S, pH 7.2, 37°C
2
anthranilate
60°C, pH 7.5, 0.05 mM Mg2+, mutant enzyme H107A/P178A
2
anthranilate
mutant D83G, in the presence of 2 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
2.7
anthranilate
mutant enzyme I36E/M47D/D83G/F149S/F193S, pH 7.2, 37°C
3.4
anthranilate
mutant enzyme I36E/M47D/D83G/F149S, pH 7.2, 37°C
3.5
anthranilate
mutant enzyme I36E/M47D/D83G/F149S/I169T, pH 7.2, 37°C
3.9
anthranilate
mutant enzyme I36E/M47D/D83G/F149S/L320M, pH 7.2, 37°C
4.2
anthranilate
60°C, pH 7.5, 0.05 mM Mg2+, wild-type enzyme
4.4
anthranilate
mutant D83G/F149S, in the presence of 0.05 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
5.1
anthranilate
60°C, pH 7.5, 0.05 mM Mg2+, mutant enzyme H107A
5.6
anthranilate
60°C, pH 7.5, 2 mM Mg2+,mutant enzyme E224Q
13.3
anthranilate
mutant D83G/F149S, in the presence of 2 mM MgCl2, 50 mM Tris-HCl, pH 7.2, at 37°C
additional information
5-phospho-alpha-D-ribose 1-diphosphate
I36E/M47D double mutant, kcat/KM: 0.0019 1/s*microM
additional information
5-phospho-alpha-D-ribose 1-diphosphate
-
I36E/M47D double mutant, kcat/KM: 0.0019 1/s*microM
additional information
5-phospho-alpha-D-ribose 1-diphosphate
wild-type, kcat/KM: 0.0023 1/s*microM
additional information
5-phospho-alpha-D-ribose 1-diphosphate
-
wild-type, kcat/KM: 0.0023 1/s*microM
additional information
anthranilate
I36E/M47D double mutant, kcat/KM: 4.8 1/s*microM
additional information
anthranilate
-
I36E/M47D double mutant, kcat/KM: 4.8 1/s*microM
additional information
anthranilate
wild-type, kcat/KM: 4.8 1/s*microM
additional information
anthranilate
-
wild-type, kcat/KM: 4.8 1/s*microM
additional information
additional information
single mutants I36E and M47D show the same kcat as I36E/M47D double mutant within experimental error
-
additional information
additional information
-
single mutants I36E and M47D show the same kcat as I36E/M47D double mutant within experimental error
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.84
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/T77I/D83G/F149S/I169T/F193S/L320M, pH 7.2, 37°C
24
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/D83G/N109S/F149S, pH 7.2, 37°C
24
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/T77I/D83G/F149S, pH 7.2, 37°C
25
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/T77I/D83G/F149S/N109S/I169T/F193S/L320M, pH 7.2, 37°C
56
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/D83G/F149S/I169T, pH 7.2, 37°C
72
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/D83G/F149S/F193S, pH 7.2, 37°C
72
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/D83G/F149S/L320M, pH 7.2, 37°C
76
5-phospho-alpha-D-ribose 1-diphosphate
mutant enzyme I36E/M47D/D83G/F149S, pH 7.2, 37°C
0.56
anthranilate
mutant enzyme I36E/M47D/T77I/D83G/F149S/I169T/F193S/L320M, pH 7.2, 37°C
13
anthranilate
mutant enzyme I36E/M47D/D83G/N109S/F149S, pH 7.2, 37°C
23
anthranilate
mutant enzyme I36E/M47D/T77I/D83G/F149S/N109S/I169T/F193S/L320M, pH 7.2, 37°C
92
anthranilate
mutant enzyme I36E/M47D/D83G/F149S/F193S, pH 7.2, 37°C
330
anthranilate
mutant enzyme I36E/M47D/D83G/F149S, pH 7.2, 37°C
360
anthranilate
mutant enzyme I36E/M47D/T77I/D83G/F149S, pH 7.2, 37°C
460
anthranilate
mutant enzyme I36E/M47D/D83G/F149S/L320M, pH 7.2, 37°C
540
anthranilate
mutant enzyme I36E/M47D/D83G/F149S/I169T, pH 7.2, 37°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
catalyzes the third step in tryptophan biosynthesis
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
32400
double mutant M47D/I36E, analytical-gel filtration chromatography
34100
double mutant M47D/I36E, analytical ultracentrifugation (sedimentation equilibrium)
60100
mutant M47D, analytical ultracentrifugation (sedimentation equilibrium)
61700
mutant M47D, analytical gel filtration chromatography
63400
mutant I36E, analytical gel filtration chromatography
66000
mutant I36E, analytical ultracentrifugation (sedimentation equilibrium)
72200
wild-type, analytical ultracentrifugation (sedimentation equilibrium)
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
monomer
double mutant M47D/I36E, analytical gel filtration chromatography and analytical ultracentrifugation (sedimentation equilibrium), Ile36 and Met47 situated at N-terminus and C-terminus of helix 3 of the small alpha-helical domain and involved in intimate intersubunit interactions
monomer or dimer
equilibrium between minor, thermo-labile monomeric and major, thermo-stable dimeric state of single mutants M47D (dissociation constant, KD: 17 +/-10 microM) and I36E (KD: 0.8 +/-0.6 microM), analytical ultracentrifugation (sedimentation equilibrium)
dimer
wild-type and mutants I36E and M47D, analytical gel filtration chromatography and analytical ultracentrifugation (sedimentation equilibrium), dimerisation mediated by residues of mainly alpha helices 1, 3 and 8 of the small alpha-helical domains (one in each subunit) in head-to-head fashion
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structures of the wild-type enzyme complexed to its two natural substrates anthranilate and 5-phosphoribosyl-1-pyrophosphate/Mg2+
hanging-drop method, crystals of ssTrpD diffract to better than 2.6 A resolution
mutant D83G/F149S in complex with 5-phospho-alpha-D-ribose 1-diphosphate and Mn2+, to 2.25 A resolution. Protein backbone of mutant D83G/F149S shows no detectable differences to the wild-type enzyme, whereas 5-phospho-alpha-D-ribose 1-diphosphate bound to mutant D83G/F149S adopts an extended conformation that contrasts markedly with the S compact shape observed in complexes of the wild-type enzyme
mutant M47D, structurally very similar to wild-type (rms deviation of 0.7 A for most of equivalent C(alpha) atoms) but reduced buried surface area per subunit compared to wild-type homodimer, Aps47 protonated at pH 6, crystals of space group P2 with four molecules (two homodimers) per asymmetric unit and A2 pseudo-symmetry, unit cell parameters a=91.6 A, b=65.9 A, c=115.7 A, beta=107.4°, 45% (v/v) solvent content, hanging drop method: 1 microlitre protein solution (5 mg/ml) + 1 microlitre reservior solution (50 mM MES pH 6.0, 18% (v/v) PEG, 5% (v/v) glycerol), room temperature, 72 h
the crystal structure of the dimeric class III phosphoribosyltransferase. The active site of this enzyme is located within the flexible hinge region of its two-domain structure. The pyrophosphate moiety of phosphoribosylpyrophosphate is coordinated by a metal ion and is bound by two conserved loop regions within this hinge region. With the structure of AnPRT available, structural analysis of all enzymatic activities of the tryptophan biosynthesis pathway is complete, thereby connecting the evolution of its enzyme members to the general development of metabolic processes. Its structure reveals it to have the same fold, topology, active site location and type of association as class II nucleoside phosphorylases. At the level of sequences, this relationship is mirrored by 13 structurally invariant residues common to both enzyme families
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D223N
kcat is 5.5fold higher than wild-type value at 2 mM Mg2+
D83G
inhibition by Mg2+ only at very high concentrations, alters the binding mode of the substrate Mg2+-5-phospho-alpha-D-ribose 1-diphosphate
D83G/F149S
E224Q
kcat is 5fold higher than wild-type value at 2 mM Mg2+
F149S
is inhibited by MgCl2 to a similar extent as wild-type, facilitates product release by increasing the conformational flexibility of the enzyme
H107A
kcat is 1.2fold higher than wild-type value at 0.05 mM Mg2+
H107A/P178A
kcat is 2.1fold lower than wild-type value at 0.05 mM Mg2+
I36E
weakened intersubunit interaction and increased protein solubility by introduction of negative side chain, monomer-dimer equilibrium (dissociation constant, KD: 0.8 +/-0.6 microM), concentration-dependent kinetic stability during heat inactivation (80°C) with half-lives from 3 to 40 min, no suitable crystal formed
I36E/M47D
mutation leads to monomerization, apparent melting temperature is 11.5°C lower than the wild-type value
I36E/M47D/D83G/F149S
mutation leads to monomerization, apparent melting temperature is 21.4°C lower than the wild-type value. kcat/Km for anthranilate is 14.5fold lower than kcat/Km for wild-type enzyme. kcat/Km for anthranilate is lower than kcat/Km for wild-type enzyme. kcat/Km for 5-phospho-alpha-D-ribose 1-diphosphate is 33fold higher than kcat/Km for wild-type enzyme
I36E/M47D/D83G/F149S/F193S
mutation leads to monomerization, apparent melting temperature is 17.4°C lower than the wild-type value. kcat/Km for anthranilate is 52fold lower than kcat/Km for wild-type enzyme. kcat/Km for anthranilate is lower than kcat/Km for wild-type enzyme. kcat/Km for 5-phospho-alpha-D-ribose 1-diphosphate is 31.3fold higher than kcat/Km for wild-type enzyme
I36E/M47D/D83G/F149S/I169T
mutation leads to monomerization, apparent melting temperature is 20.6°C lower than the wild-type value. kcat/Km for anthranilate is 8.9fold lower than kcat/Km for wild-type enzyme. kcat/Km for anthranilate is lower than kcat/Km for wild-type enzyme. kcat/Km for 5-phospho-alpha-D-ribose 1-diphosphate is 24.3fold higher than kcat/Km for wild-type enzyme
I36E/M47D/D83G/F149S/L320M
mutation leads to monomerization, apparent melting temperature is 20.9°C lower than the wild-type value. kcat/Km for anthranilate is 11fold lower than kcat/Km for wild-type enzyme. kcat/Km for anthranilate is lower than kcat/Km for wild-type enzyme. kcat/Km for 5-phospho-alpha-D-ribose 1-diphosphate is 31.3fold higher than kcat/Km for wild-type enzyme
I36E/M47D/D83G/N109S/F149S
mutation leads to monomerization, apparent melting temperature is 20.5°C lower than the wild-type value. kcat/Km for anthranilate is 39fold lower than kcat/Km for wild-type enzyme. kcat/Km for anthranilate is lower than kcat/Km for wild-type enzyme. kcat/Km for 5-phospho-alpha-D-ribose 1-diphosphate is 10fold higher than kcat/Km for wild-type enzyme
I36E/M47D/D83G/N109S/F149S/I169T/L320M/N324I
mutation leads to monomerization, apparent melting temperature is 18.5°C lower than the wild-type value
I36E/M47D/T77I/D83G/F149S
mutation leads to monomerization, apparent melting temperature is 13.3°C lower than the wild-type value. kcat/Km for anthranilate is 13.3fold lower than kcat/Km for wild-type enzyme. kcat/Km for anthranilate is lower than kcat/Km for wild-type enzyme. kcat/Km for 5-phospho-alpha-D-ribose 1-diphosphate is 10fold higher than kcat/Km for wild-type enzyme
I36E/M47D/T77I/D83G/F149S/F193S
mutation leads to monomerization, apparent melting temperature is 11.4°C lower than the wild-type value
I36E/M47D/T77I/D83G/F149S/I169T/F193S/L320M
mutation leads to monomerization, apparent melting temperature is 9.1°C lower than the wild-type value. kcat/Km for anthranilate is 85.7fold lower than kcat/Km for wild-type enzyme. kcat/Km for anthranilate is lower than kcat/Km for wild-type enzyme. kcat/Km for 5-phospho-alpha-D-ribose 1-diphosphate is 2.7fold lower than kcat/Km for wild-type enzyme
I36E/M47D/T77I/D83G/F149S/N109S/I169T/F193S/L320M
mutation leads to monomerization, apparent melting temperature is 8.7°C lower than the wild-type value. kcat/Km for anthranilate is 209fold lower than kcat/Km for wild-type enzyme. kcat/Km for anthranilate is lower than kcat/Km for wild-type enzyme. kcat/Km for 5-phospho-alpha-D-ribose 1-diphosphate is higher than kcat/Km for wild-type enzyme
K106Q
kcat is 2.3fold lower than wild-type value at 2 mM Mg2+
M47D
weakened intersubunit interaction and increased protein solubility by introduction of negative side chain, monomer-dimer equilibrium (dissociation constant, KD: 17 +/-10 microM), concentration-dependent kinetic stability during heat inactivation (80°C) with half-lives from 4 to 15 min, no structural perturbation
M47D/I36E
double mutant, monomeric, similar catalytic efficiencies as the wild-type for both substrates, first-order kinetics for time-dependent but not concentration-dependent heat inactivation at 80°C with half-live t1/2: 3 min, no suitable crystal formed
R164A
kcat is 6.8fold lower than wild-type value at 0.05 mM Mg2+
R164A/H154A
kcat is 5.8fold lower than wild-type value at 0.05 mM Mg2+
D83G/F149S
activity increases up to about 2-5 mM MgCl2 and decreases only moderately at higher Mg2+ concentrations
D83G/F149S
mutation leads to monomerization, apparent melting temperature is 9.5°C lower than the wild-type value
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
70
t1/2: 69 min, addition of anthranilate has no stabilizing effect
80
half-lives (t1/2) as measure of kinetic stability, t1/2(wild-type): 40 min, t1/2(double mutant M47D/I36E): 3 min, t1/2(mutant I36E, about 1 microM): 3 min, t1/2(mutant I36E, about 20 microM): 40 min, t1/2(mutant M47D, about 1 microM): 4 min, t1/2(mutant M47D, about 47 microM): 15 min, irreversible heat inactivation (pH 6.7) at different time points followed by chilling, centrifugation and estimation of residual activity
82
double mutant I36E/M47D, 12 microM, melting temperature at which half of the protein is unfolded deduced from DSC, pH 7.5
83
mutant M47D, 12 microM, melting temperature at which half of the protein is unfolded deduced from DSC, pH 7.5
85
t1/2: 35 min, addition of anthranilate has no stabilizing effect
90
92
wild-type, 12 microM, melting temperature at which half of the protein is unfolded deduced from differential scanning calorimetry (DSC), pH 7.5
additional information
90
mutant I36E, 12 microM, melting temperature at which half of the protein is unfolded deduced from DSC, pH 7.5
90
wild-type protein and mutant D83G show melting temperatures of 90 and 91°C, respectively, mutant F149S protein and the double mutant D83G/F149S exhibit melting temperature values of 82°C
additional information
concentration-dependent kinetic stability of single mutants I36E and M47D during heat inactivation at 80°C
additional information
-
concentration-dependent kinetic stability of single mutants I36E and M47D during heat inactivation at 80°C
additional information
dimerisation stabilizes against thermal denaturation
additional information
-
dimerisation stabilizes against thermal denaturation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
the enzyme is either digested by trypsin, V8-protease or thermolysin after incubation for 1 h at 25°C but not by thermolysin at 70°C
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
from Escherichia coli extract by heat-precipitation of host proteins followed by metal chelate affinity chromatography, yield of 0.4-0.8 mg protein per 1 g wet cell mass, >95% purity, stored at -80°C
wild-type and mutants purified by heat precipitation and metal chelate affinity chromatography
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
amplified fragments containing the full-length trpD gene ligated into the pQE40 vector, transformed into Escherichia coli competent DH5alpha cells. Wild-type and its mutants expressed heterologously in Escherichia coli strain W3110 trpEA2trpEA2, containing the helper plasmid pDM,1
wild-type and mutants in pQE40 for expression with N-terminal hexa-His tag in Escherichia coli W3110 trpEA2 (pDM)
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Ivens, A.; Mayans, O.; Szadkowski, H.; Wilmanns, M.; Kirschner, K.
Purification, characterization and crystallization of thermostable anthranilate phosphoribosyltransferase from Sulfolobus solfataricus
Eur. J. Biochem.
268
2246-2252
2001
Saccharolobus solfataricus, Saccharolobus solfataricus (P50384)
Marino, M.; Deuss, M.; Svergun, D.I.; Konarev, P.V.; Sterner, R.; Mayans, O.
Structural and mutational analysis of substrate complexation by anthranilate phosphoribosyltransferase from Sulfolobus solfataricus
J. Biol. Chem.
281
21410-21421
2006
Saccharolobus solfataricus (P50384), Saccharolobus solfataricus
Schwab, T.; Skegro, D.; Mayans, O.; Sterner, R.
A rationally designed monomeric variant of anthranilate phosphoribosyltransferase from Sulfolobus solfataricus is as active as the dimeric wild-type enzyme but less thermostable
J. Mol. Biol.
376
506-516
2008
Saccharolobus solfataricus (P50384), Saccharolobus solfataricus
Schlee, S.; Deuss, M.; Bruning, M.; Ivens, A.; Schwab, T.; Hellmann, N.; Mayans, O.; Sterner, R.
Activation of anthranilate phosphoribosyltransferase from Sulfolobus solfataricus by removal of magnesium inhibition and acceleration of product release
Biochemistry
48
5199-5209
2009
Saccharolobus solfataricus (P50384), Saccharolobus solfataricus
Schwab, T.; Sterner, R.
Stabilization of a metabolic enzyme by library selection in Thermus thermophilus
Chembiochem
12
1581-1588
2011
Saccharolobus solfataricus (P50384), Saccharolobus solfataricus, Saccharolobus solfataricus P2 (P50384)
Mayans, O.; Ivens, A.; Nissen, L.J.; Kirschner, K.; Wilmanns, M.
Structural analysis of two enzymes catalysing reverse metabolic reactions implies common ancestry
EMBO J.
21
3245-3254
2002
Saccharolobus solfataricus (P50384), Saccharolobus solfataricus P2 (P50384)
Schlee, S.; Straub, K.; Schwab, T.; Kinateder, T.; Merkl, R.; Sterner, R.
Prediction of quaternary structure by analysis of hot spot residues in protein-protein interfaces the case of anthranilate phosphoribosyltransferases
Proteins
87
815-825
2019
Acetomicrobium mobile, Coprococcus eutactus, Fructobacillus fructosus, Methanocella conradii, Methanococcus voltae, Pelodictyon luteolum, Pelodictyon luteolum DSM270, Petrotoga mobilis, Saccharolobus solfataricus (P50384), Saccharolobus solfataricus P2 (P50384), Staphylococcus aureus, Staphylococcus aureus USA300, Staphylococcus haemolyticus, Staphylococcus haemolyticus JCSC1435
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