BRENDA - Enzyme Database

The Structure of Rauvolfia serpentina Strictosidine Synthase Is a Novel Six-Bladed {beta}-Propeller Fold in Plant Proteins

Ma, X.; Panjikar, S.; Koepke, J.; Loris, E.; Stockigt, J.; Plant Cell 18, 907-920 (2006)

Data extracted from this reference:

Application
EC Number
Application
Commentary
Organism
4.3.3.2
medicine
anti-arrhythmic heart disorders, ajmaline agents
Rauvolfia serpentina
4.3.3.2
synthesis
involved in the biosynthesis of almost all plant monoterpenoid indole alkaloids
Rauvolfia serpentina
Cloned(Commentary)
EC Number
Commentary
Organism
4.3.3.2
cloned with His-tag, expression in Escherichia coli strain M15, after expression and Ni-NTA column removed by DAPase
Rauvolfia serpentina
Crystallization (Commentary)
EC Number
Crystallization
Organism
4.3.3.2
crystals of free STR1 and co-crystallization of STR1 with each tryptamine and secologanin, crystal growing by hanging drop vapor diffusion, 302-304 K, 0.8 M potassium sodium tartrate tetrahydrate, pH 7.5 - 8.0, combined multiple wavelength anomalous dispersion and molecular replacement methods
Rauvolfia serpentina
Engineering
EC Number
Amino acid exchange
Commentary
Organism
4.3.3.2
E309A
mutant with 25% decrease of kcat
Rauvolfia serpentina
4.3.3.2
H307A
mutant with 130-fold increase of Km for secologanin
Rauvolfia serpentina
4.3.3.2
Y151F
mutant with 2.8-fold increase of Km for tryptamin
Rauvolfia serpentina
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
4.3.3.2
2,4-dinitrofluorobenzene
5 mM, no inactivation after 30 min and 15% inactivation after 120 min in Tris-HCl, pH 8.0
Rauvolfia serpentina
4.3.3.2
4-(2-aminoethyl)-benzenesulfonyl fluoride
4 mM, 4% inactivation after 30 min and 12% inactivation after 120 min in 100 mM potassium phosphate buffer, pH 7.0
Rauvolfia serpentina
4.3.3.2
Diethylpyrocarbonate
1 mM, 49% inactivation after 30 min and 63% inactivation after 120 min in 100 mM potassium phosphate, pH 6.0
Rauvolfia serpentina
4.3.3.2
additional information
no inactivation by phenylmethanesulfonyl fluoride, L-chloro-3-(4-tosyl-amido)-4-phenyl-2-butanone and L-chloro-3-(4-tosyl-amido)-7-amino-2-heptanone after 30 and 120 min in 100 mM potassium phosphate buffer, pH 7.0
Rauvolfia serpentina
4.3.3.2
N,N'-dicyclohexylcarbodiimide
0.05 mM, complete inactivation after 30 min in MES, pH 6.0
Rauvolfia serpentina
4.3.3.2
N-Acetylimidazole
10 mM, 5% inactivation after 30 min and 12% inactivation after 120 min in Tris-HCl, pH 8.0
Rauvolfia serpentina
4.3.3.2
N-ethyl-5-phenylisoxazolium-3'-sulfonate
1 mM leads to 48% inactivation after 30 min and 53% inactivation after 120 min in MES, pH 6.0, 5 mM leads to 80% inactivation after 30 min and 100% inactivation after 120 min in MES, pH 6.0
Rauvolfia serpentina
4.3.3.2
p-chlormercuribenzoate
1 mM, 34% inactivation after 30 min and 35% inactivation after 120 min in 100 mM potassium phosphate, pH 6.0
Rauvolfia serpentina
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.3.2
0.0054
-
tryptamine
E309A, pH 7.0, 35°C, 15 min
Rauvolfia serpentina
4.3.3.2
0.0062
-
tryptamine
wild type, pH 7.0, 35°C, 15 min
Rauvolfia serpentina
4.3.3.2
0.0079
-
tryptamine
H307A, pH 7.0, 35°C, 15 min
Rauvolfia serpentina
4.3.3.2
0.0172
-
tryptamine
Y151F, pH 7.0, 35°C, 15 min
Rauvolfia serpentina
4.3.3.2
0.039
-
secologanin
wild type, pH 7.0, 35°C, 15 min
Rauvolfia serpentina
4.3.3.2
0.044
-
secologanin
Y151F, pH 7.0, 35°C, 15 min
Rauvolfia serpentina
4.3.3.2
0.095
-
secologanin
E309A, pH 7.0, 35°C, 15 min
Rauvolfia serpentina
4.3.3.2
5.07
-
secologanin
H307A, pH 7.0, 35°C, 15 min
Rauvolfia serpentina
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
4.3.3.2
Rauvolfia serpentina
P68175
-
-
Purification (Commentary)
EC Number
Commentary
Organism
4.3.3.2
-
Rauvolfia serpentina
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.3.3.2
5,6-dihydroxytryptamine
-
666582
Rauvolfia serpentina
?
-
-
-
?
4.3.3.2
5-fluorotryptamine
-
666582
Rauvolfia serpentina
?
-
-
-
?
4.3.3.2
6-fluorotryptamine
-
666582
Rauvolfia serpentina
?
-
-
-
?
4.3.3.2
6-methoxytryptamine
-
666582
Rauvolfia serpentina
?
-
-
-
?
4.3.3.2
7-methyltryptamine
-
666582
Rauvolfia serpentina
?
-
-
-
?
4.3.3.2
serotonin
-
666582
Rauvolfia serpentina
?
-
-
-
?
4.3.3.2
tryptamine + secologanin
-
666582
Rauvolfia serpentina
3alpha(S)-strictosidine
-
-
-
?
Application (protein specific)
EC Number
Application
Commentary
Organism
4.3.3.2
medicine
anti-arrhythmic heart disorders, ajmaline agents
Rauvolfia serpentina
4.3.3.2
synthesis
involved in the biosynthesis of almost all plant monoterpenoid indole alkaloids
Rauvolfia serpentina
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
4.3.3.2
cloned with His-tag, expression in Escherichia coli strain M15, after expression and Ni-NTA column removed by DAPase
Rauvolfia serpentina
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
4.3.3.2
crystals of free STR1 and co-crystallization of STR1 with each tryptamine and secologanin, crystal growing by hanging drop vapor diffusion, 302-304 K, 0.8 M potassium sodium tartrate tetrahydrate, pH 7.5 - 8.0, combined multiple wavelength anomalous dispersion and molecular replacement methods
Rauvolfia serpentina
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
4.3.3.2
E309A
mutant with 25% decrease of kcat
Rauvolfia serpentina
4.3.3.2
H307A
mutant with 130-fold increase of Km for secologanin
Rauvolfia serpentina
4.3.3.2
Y151F
mutant with 2.8-fold increase of Km for tryptamin
Rauvolfia serpentina
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
4.3.3.2
2,4-dinitrofluorobenzene
5 mM, no inactivation after 30 min and 15% inactivation after 120 min in Tris-HCl, pH 8.0
Rauvolfia serpentina
4.3.3.2
4-(2-aminoethyl)-benzenesulfonyl fluoride
4 mM, 4% inactivation after 30 min and 12% inactivation after 120 min in 100 mM potassium phosphate buffer, pH 7.0
Rauvolfia serpentina
4.3.3.2
Diethylpyrocarbonate
1 mM, 49% inactivation after 30 min and 63% inactivation after 120 min in 100 mM potassium phosphate, pH 6.0
Rauvolfia serpentina
4.3.3.2
additional information
no inactivation by phenylmethanesulfonyl fluoride, L-chloro-3-(4-tosyl-amido)-4-phenyl-2-butanone and L-chloro-3-(4-tosyl-amido)-7-amino-2-heptanone after 30 and 120 min in 100 mM potassium phosphate buffer, pH 7.0
Rauvolfia serpentina
4.3.3.2
N,N'-dicyclohexylcarbodiimide
0.05 mM, complete inactivation after 30 min in MES, pH 6.0
Rauvolfia serpentina
4.3.3.2
N-Acetylimidazole
10 mM, 5% inactivation after 30 min and 12% inactivation after 120 min in Tris-HCl, pH 8.0
Rauvolfia serpentina
4.3.3.2
N-ethyl-5-phenylisoxazolium-3'-sulfonate
1 mM leads to 48% inactivation after 30 min and 53% inactivation after 120 min in MES, pH 6.0, 5 mM leads to 80% inactivation after 30 min and 100% inactivation after 120 min in MES, pH 6.0
Rauvolfia serpentina
4.3.3.2
p-chlormercuribenzoate
1 mM, 34% inactivation after 30 min and 35% inactivation after 120 min in 100 mM potassium phosphate, pH 6.0
Rauvolfia serpentina
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
4.3.3.2
0.0054
-
tryptamine
E309A, pH 7.0, 35°C, 15 min
Rauvolfia serpentina
4.3.3.2
0.0062
-
tryptamine
wild type, pH 7.0, 35°C, 15 min
Rauvolfia serpentina
4.3.3.2
0.0079
-
tryptamine
H307A, pH 7.0, 35°C, 15 min
Rauvolfia serpentina
4.3.3.2
0.0172
-
tryptamine
Y151F, pH 7.0, 35°C, 15 min
Rauvolfia serpentina
4.3.3.2
0.039
-
secologanin
wild type, pH 7.0, 35°C, 15 min
Rauvolfia serpentina
4.3.3.2
0.044
-
secologanin
Y151F, pH 7.0, 35°C, 15 min
Rauvolfia serpentina
4.3.3.2
0.095
-
secologanin
E309A, pH 7.0, 35°C, 15 min
Rauvolfia serpentina
4.3.3.2
5.07
-
secologanin
H307A, pH 7.0, 35°C, 15 min
Rauvolfia serpentina
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
4.3.3.2
-
Rauvolfia serpentina
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
4.3.3.2
5,6-dihydroxytryptamine
-
666582
Rauvolfia serpentina
?
-
-
-
?
4.3.3.2
5-fluorotryptamine
-
666582
Rauvolfia serpentina
?
-
-
-
?
4.3.3.2
6-fluorotryptamine
-
666582
Rauvolfia serpentina
?
-
-
-
?
4.3.3.2
6-methoxytryptamine
-
666582
Rauvolfia serpentina
?
-
-
-
?
4.3.3.2
7-methyltryptamine
-
666582
Rauvolfia serpentina
?
-
-
-
?
4.3.3.2
serotonin
-
666582
Rauvolfia serpentina
?
-
-
-
?
4.3.3.2
tryptamine + secologanin
-
666582
Rauvolfia serpentina
3alpha(S)-strictosidine
-
-
-
?