Information on EC 4.3.2.1 - argininosuccinate lyase

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

EC NUMBER
COMMENTARY hide
4.3.2.1
-
RECOMMENDED NAME
GeneOntology No.
argininosuccinate lyase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
2-(Nomega-L-arginino)succinate = fumarate + L-arginine
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
elimination
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Alanine, aspartate and glutamate metabolism
-
-
Arginine biosynthesis
-
-
arginine metabolism
-
-
Biosynthesis of antibiotics
-
-
Biosynthesis of secondary metabolites
-
-
canavanine biosynthesis
-
-
L-arginine biosynthesis I (via L-ornithine)
-
-
L-arginine biosynthesis II (acetyl cycle)
-
-
L-arginine biosynthesis III (via N-acetyl-L-citrulline)
-
-
L-arginine biosynthesis IV (archaebacteria)
-
-
L-citrulline-nitric oxide cycle
-
-
Metabolic pathways
-
-
urea cycle
SYSTEMATIC NAME
IUBMB Comments
2-(Nomega-L-arginino)succinate arginine-lyase (fumarate-forming)
-
CAS REGISTRY NUMBER
COMMENTARY hide
9027-34-3
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
subspecies Lactobacillus plantarum plantarum
-
-
Manually annotated by BRENDA team
Mus musculus BALB/c
gene asl
-
-
Manually annotated by BRENDA team
gene Rv1659
UniProt
Manually annotated by BRENDA team
gene Rv1659
UniProt
Manually annotated by BRENDA team
strain PCC 73102
SwissProt
Manually annotated by BRENDA team
Gulf toadfish
-
-
Manually annotated by BRENDA team
cv. Nipponbare, gene OsASL1
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(Nomega-L-arginino)succinate
fumarate + L-arginine
show the reaction diagram
2-(Nomega-L-arginino)succinate
fumarate + L-arginine
show the reaction diagram
arginine + fumarate
argininosuccinate
show the reaction diagram
-
-
-
-
r
argininosuccinate
?
show the reaction diagram
-
urea cycle enzyme, deficiency results in argininosuccinic aciduria
-
-
-
argininosuccinate
fumarate + L-Arg
show the reaction diagram
difluorofumarate + L-Arg
?
show the reaction diagram
-
-
-
-
?
fumarate + arginine
argininosuccinate
show the reaction diagram
-
-
-
-
r
fumarate + L-Arg
argininosuccinate
show the reaction diagram
fumarate + L-arginine
2-(Nomega-L-arginino)succinate
show the reaction diagram
fumarate + L-canavanine
?
show the reaction diagram
-
at 78% of the activity with L-Arg
-
-
?
monofluorofumarate + L-Arg
?
show the reaction diagram
-
-
-
-
?
omega-N-(L-arginino)succinate
L-arginine + fumarate
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(Nomega-L-arginino)succinate
fumarate + L-arginine
show the reaction diagram
2-(Nomega-L-arginino)succinate
fumarate + L-arginine
show the reaction diagram
argininosuccinate
?
show the reaction diagram
-
urea cycle enzyme, deficiency results in argininosuccinic aciduria
-
-
-
argininosuccinate
fumarate + L-Arg
show the reaction diagram
fumarate + L-arginine
2-(Nomega-L-arginino)succinate
show the reaction diagram
omega-N-(L-arginino)succinate
L-arginine + fumarate
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
GDP
-
0.2 mM, slight stimulation
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-ethyl-3-(3-diethylaminopropyl)carbodiimide
-
at pH 6
acetylenedicarboxylate
-
-
Arg
-
noncompetitive versus argininosuccinate
argininosuccinate
-
0.3 mM inhibits the reaction by 97%
bromomesaconic acid
-
argininosuccinate completely protects, Lys51 has an essential role in binding of argininosuccinate and consequently is essential in catalysis; inactivation follows pseudo-first-order kinetics
citrulline
D-glucose
-
12.5 mM glucose decreases activity of ASL by 20%, 25 mM glucose by 50%
diethyl dicarbonate
difluorofumarate
-
pseudo-first-order process
fumarate
-
noncompetitive versus argininosuccinate
monofluorofumarate
-
reversible, pseudo-first-order process
N3-(L-1-carboxy-2-nitroethyl)-L-arginine
-
-
succinate
Urea
-
competitive
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
amino acids
-
extra amino acids in culture media increases activity of ASL by decreasing inhibitory acetylation rate
GTP
-
0.2 mM, stimulates
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.2 - 2.6
(Nomega-L-arginino)succinate
0.007 - 1.7
argininosuccinate
0.046
difluorofumarate
-
-
0.045 - 3.159
disodium argininosuccinate
0.067 - 5.3
fumarate
1.4
monofluorofumarate
-
-
0.12
N,N'-bis(tert-butoxycarbonyl)-5-[(3-aminopropyl)amino]pentan-1-ol
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.12 - 2930
argininosuccinate
34.5
N,N'-bis(tert-butoxycarbonyl)-5-[(3-aminopropyl)amino]pentan-1-ol
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0012
-
activity in lung homogenate
0.006
-
pH and temperature not specified in the publication, activity in cell culture in minimal growth medium with 20 mM glucose and 5 mM NH4+
3
-
mutant c.C1153T, pH 7.5, 37C
5
-
mutant c.A943G, pH 7.5, 37C
8.6
-
pRH1, units/mg
41
-
mutant c.A566G, pH 7.5, 37C
208
-
mutant c.G532A, pH 7.5, 37C
280
-
mutant c.C1135T, pH 7.5, 37C
4227
-
wild-type, pH 7.5, 37C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.8 - 7.2
-
formation of argininosuccinate
7.4
-
activity assay
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
activity assay
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.2
-
isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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present in neurones and nerve fibres in the myenteric plexus of the lower oesophagael sphincter, antrum, pylorus, ileum and colon, in the submucosal plexus of ileum and colon, in longitudinal muscle of ileum and colon, nerve bundles of circular muscles
Manually annotated by BRENDA team
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ASL expression is time-dependently reduced in hypoxia
Manually annotated by BRENDA team
using DNA
Manually annotated by BRENDA team
-
expression of enzyme genetic variant OsASL1.1
Manually annotated by BRENDA team
-
expression of enzyme genetic variant OsASL1.1
Manually annotated by BRENDA team
-
low mRNA content
Manually annotated by BRENDA team
-
nerve fibers of smooth muscle layer
Manually annotated by BRENDA team
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fumarate hydratase defiecient or proficient cells, the latter termed UOKpFH cells
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
expression of OsASL1.1
Manually annotated by BRENDA team
-
expression of OsASL1.1
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
50000
-
SDS-PAGE
51700
-
SDS-PAGE
52340
the asl open reading frame encodes a protein of 464 amino acids with a calculated mass of 52337 Da
187000
-
equilibrium sedimentation
190000
200000
202000
-
gel filtration
215000
native PAGE, recombinant ASL
218000
-
gel filtration
220000
gel filtration, recombinant fusion protein
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
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5 or 6 * 39000, SDS-PAGE
homotetramer
pentamer
-
5 or 6 * 39000, SDS-PAGE
tetramer
additional information
-
small heat shock protein, alphaA-crystallin, functions as a molecular chaperone, and enhances thermal stability of both delta-crystallin and ASL. Thermal unfolding of delta-crystallin or ASL in the presence of alphaA-crystallin follows a similar three-state model. A stable intermediate which retains about 30% alpha-helical structure is observed. Protection from thermal denaturation by alphaA-crystallin is by interaction with partly unfolded ASL to form high molecular weight heteroligomers. Aggregate formation of ASL is significantly reduced in the presence of alphaA-crystallin. The extent of protection of ASL at different ratios of alpahA-crystallin is described by hyperbolic curves, suggesting the preferential recognition of partly unfolded ASL by alphaA-crystallin
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
side-chain modification
-
acetylation at Lys288 or Lys69 in liver decreases activity of ASL
additional information
-
origin of the multiple forms is due to post-translational oxidative modification of the protein
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
S283A mutant, hanging-drop vapor diffusion at room temperatur, 9 mg/ml protein in 10 mM Tris-HCl, pH 7.5 and 1 mM EDTA is preincubated with 75 mM argininosuccinate, subsequently 0.005 ml drops of the solution are mixed with an equal ammount of precipitating solution consisting of 12% polyethylene glycol 2000 MME, 300 mM MgCl2, 100 mM HEPES, pH 7.4, crystals diffract to 1.96 A resolution
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apoenzyme and complex of double loop mutant with sulfate, growing at room temperature using the hanging drop vapor diffusion method, molecular replacement at 2.2 A resolution for the DLM-sulfate complex, 2.5 A resolution for the apoenzyme
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crystallized from a highly concentrated sample of purified recombinant alpha-methylacyl-CoA-racemase with arginosuccinate lyase as a minor impurity, growing at room temperature in mother liquid consisting of 1.26 M ammonium phosphate pH 7.0, small bipyramidal crystals, molecular replacement at 2.44 A resolution
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hanging-drop vapor diffusion, crystal structure of Q286R mutant at 2.65 A
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purified recombinant His-tagged enzyme, microbatch-under-oil method, mixing of 0.002 ml of 9.5 mg/ml protein in 25 mM phosphate buffer, pH 7.4, with 0.002 ml of precipitant solution containing 100 mM Bis-Tris, pH 5.5, 25% w/v polyethylene glycol 3350, X-ray diffraction structure determination and analysis at 2.40 A resolution, molecular replacement method
to 2.5 A resolution, space group R3
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 8.5
-
stable for at least 20 min
34436
7
-
at 4C and at 25C, 8 h, stable
34429
8.2
-
25C, about 40% loss of activity after 8 h; 4C, about 60% loss of activity after 4 h
34429
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
Tris-HCl buffer, pH 8.5, 50% loss of activity after 2.3 h
38
-
30 min, stable
49
-
20 min, 50% inactivation
52
-
20 min, 50% inactivation
54
-
mid-point temperature transition 2
57
-
mid-point temperature transition 1 and 3
58
-
mid-point temperature transition 3, presence of alphaA-crystallin
65
-
complete denaturation
83
-
mid-point temperature transition 1, presence of alphaA-crystallin
85
-
mid-point temperature transition 2, presence of alphaA-crystallin
additional information
-
enzyme forms with higher isoelectric point are much more thermostable than those with lower pI values
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
Arg or phosphate ions protect from cold inactivation
-
ASL undergoes cold dissociation via a dimer intermediate
-
phosphate, arsenate, 20% dimethylsulfoxide or 20% glycerol protects against cold inactivation
-
repeated freezing and thawing does not appreciably alter the activity
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
0C, Tris buffer, pH 8.5, 50% loss of activity after 2.5 h
-
4C, Tris-HCl buffer, pH 8.5, 50% loss of activity after 2.3 h
-
frozen in 0.1 M potassium phosphate, pH 7.0, stable
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
brain
-
kidney
-
leaf extract is centrifuged, the supernatant lyophilized, the resuspended material is partially purified on a G-15 Sephadex and a Q-Sepharose FF column
-
liver ASL
-
recombinant ASL
-
recombinant C-terminally His-tagged enzyme from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography and dialysis to homogeneity
recombinant His-tagged ASL
recombinant protein
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning of argH gene encoding ASL
-
cloning of cDNA
-
expressed as His tag enzyme
-
expressed as His tag enzymes
-
expression in Escherichia coli
expression in Escherichia coli and COS-7 cells
-
expression in Escherichia coli argH auxotroph, deficient in argininosuccinate lyase
-
expression in mouse cells
-
expression in Saccharomyces cerevisiae and in Escherichia coli
-
gene argH, expression of Rv1659 as His-tagged protein, with an extra methionine at the N-terminus and an extended sequence LEHHHHHH at the C-terminus, in Escherichia coli strain BL21 (DE3)
gene asl, quantitative realtime PCR expression analysis in infected and control cells
-
gene OsASL1, two variants OsASL1.1 and OsASL1.2 resulting from different transcription start sites, map-based cloning, only OsASL1.1 is able to complement the ref1 mutant phenotype, functional expression of OsASL1.1 in Saccharomyces cerevisiae, plastid-localized expression of OsASL1.1-GFP fusion protein in onion epidermal cells
-
in vitro overexpression of ASL enzyme mutant R236W in COS7 cells andprimary fibroblasts
-
into the pGEM-T vector for sequence analysis
overexpressed in HEK293T cells or in Escherichia coli
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
Brucella abortus strain 2308 infection highly induces enzyme expression in RAW-264.7 cells
expression of ASL is significantly decreased to 50% in Shunt lambs
-
mRNA levels in liver decrease significantly when gulf toadfishes are crowded by 7 days
-
threefold repression of enzyme formation by arginine
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E296Q
-
retains only 0.001% residual activity
N116L
-
no activity
R115N
-
no activity
S283A
-
no activity
S384A
-
catalytic efficiency is decreased by an order of magnitude in comparison to wild-type delta II crystallin
T161V
-
no activity
Y323F
-
catalytic efficiency is decreased by an order of magnitude in comparison to wild-type delta II crystallin
Y323I
-
no activity
A41G/I43M
-
mutant without enzyme activity
D115A
-
mutant with 45% enzyme activity
D115E
-
mutant with 42.6% enzyme activity
E115D
-
mutant without enzyme activity
I143M
-
mutant without enzyme activity
M9F/E115D
-
mutant with 3.3% enzyme activity
M9W
-
mutant with 2.0% enzyme activity
M9W/E115D
-
mutant with 17% enzyme activity
M9W/V14S
-
mutant with 1.9% enzyme activity
M9Y/E115D
-
mutant with 5.5% enzyme activity
TRUN1-8
-
mutant with 37% enzyme activity
TRUN1-9
-
mutant with 2.1% enzyme activity
W9A
-
mutant with 1.9% enzyme activity
W9F
-
mutant with 11.3% enzyme activity
W9M
-
mutant with 1.0% enzyme activity
W9R
-
mutant with 3% enzyme activity
W9Y
-
mutant with 49.5% enzyme activity
A398D
-
no activity, 15.55 of wild-type activity if coexpressed with Q286R
D31N
missense mutation found in patients with late onset of argininosuccinic aciduria, residual activity
D87G
-
no activity, 35.2% of wild-type activity if coexpressed with Q286R
E73K
missense mutation found in patient with neonatal onset of argininosuccinic aciduria, enzyme inactive
K288R
-
132% of wild-type activity
M360T
-
no actiivty, 10% of wild-type activity if coexpressed with Q286R
M382R
missense mutation found in patient with neonatal onset of argininosuccinic aciduria, residual activity
R12Q
-
6% of wild-type kcat
R182Q
missense mutation found in patient with late onset of argininosuccinic aciduria, enzyme inactive
R186Q
missense mutation found in patient with neonatal onset of argininosuccinic aciduria, residual activity
R297Q
missense mutation found in patient with late onset of argininosuccinic aciduria, residual activity
R456W missense mutation found in patient with neonatal onset of argininosuccinic aciduria
, residual activity
V178M
missense mutation combined with mutation R186Q on the other allele found in patient with neonatal onset of argininosuccinic aciduria, enzyme activity reduced compared to wild-type
V335L
missense mutation found in patients with late onset of argininosuccinic aciduria, enzyme activity reduced compared to wild-type
R140L
-
naturally occuring mutation of variant OsASL1.1, which renders the enzyme catalytically inactive
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
the rapid and sensitive assay method is used to detect increases in the activity in serum from patients with liver diseases
diagnostics
medicine
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