Information on EC 4.4.1.8 - cystathionine beta-lyase

New: Word Map on EC 4.4.1.8
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Search Reference ID:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


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

EC NUMBER
COMMENTARY hide
4.4.1.8
-
RECOMMENDED NAME
GeneOntology No.
cystathionine beta-lyase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
2-aminoprop-2-enoate = 2-iminopropanoate
show the reaction diagram
spontaneous
-
-
-
2-iminopropanoate + H2O = pyruvate + NH3
show the reaction diagram
spontaneous
-
-
-
L-cystathionine + H2O = L-homocysteine + pyruvate + NH3
show the reaction diagram
L-cystathionine = L-homocysteine + 2-aminoprop-2-enoate
show the reaction diagram
(1a)
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
elimination
-
-
of RSH, C-S bond cleavage
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
-
Cysteine and methionine metabolism
-
-
Metabolic pathways
-
-
methionine metabolism
-
-
seleno-amino acid biosynthesis
-
-
Selenocompound metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
L-cystathionine L-homocysteine-lyase (deaminating; pyruvate-forming)
A pyridoxal-phosphate protein. The enzyme cleaves a carbon-sulfur bond, releasing L-homocysteine and an unstable enamine product that tautomerizes to an imine form, which undergoes a hydrolytic deamination to form pyruvate and ammonia. The latter reaction, which can occur spontaneously, can also be catalysed by EC 3.5.99.10, 2-iminobutanoate/2-iminopropanoate deaminase. The enzyme from some sources also acts on L-cystine, forming pyruvate, ammonia and cysteine persulfide, and a number of related compounds. Possibly identical, in yeast, with EC 4.4.1.6 S-alkylcysteine lyase.
CAS REGISTRY NUMBER
COMMENTARY hide
9055-05-4
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
subsp. bulgaricus
-
-
Manually annotated by BRENDA team
strain 8944
-
-
Manually annotated by BRENDA team
strain 8944
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
FM518
-
-
Manually annotated by BRENDA team
FM518
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
L. cv. Desiree
Uniprot
Manually annotated by BRENDA team
strains AX3 and JCSC1435, gene metC or SH2636
-
-
Manually annotated by BRENDA team
strain ISP5230
SwissProt
Manually annotated by BRENDA team
strain ISP5230
SwissProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
cystathionine beta-lyase is part of the CYS3-controlled regulatory network
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
(S)-4-(4-methylpentan-2-one)-L-cysteine + H2O
4-mercapto-4-methylpentan-2-one + NH3 + pyruvate
show the reaction diagram
-
Str3p is able to release 0.0123 mM 4-mercapto-4-methylpentan-2-one from 2 mM concentration of its precursor S-(2-methyl-4-oxopentan-2-yl)-L-cysteine
-
?
cystathionine + H2O
?
show the reaction diagram
cystathionine + H2O
L-homocysteine + NH3 + pyruvate
show the reaction diagram
cystine + H2O
pyruvate + NH3 + thiocysteine
show the reaction diagram
-
-
-
-
?
D-Cys + H2O
?
show the reaction diagram
-
-
-
-
?
D-cystine + H2O
?
show the reaction diagram
-
2% of the activity with L-cystine
-
?
DL-homocysteine + H2O
hydrogen sulfide + 2-oxobutyrate + ?
show the reaction diagram
homoserine + H2O
?
show the reaction diagram
-
96% of the activity with L-cystathionine
-
-
?
L-cystathionine
L-homocysteine + pyruvate + NH4+
show the reaction diagram
L-cystathionine + H2O
cysteine + ?
show the reaction diagram
-
alpha,gamma-elimination
-
?
L-cystathionine + H2O
L-homocysteine + NH3 + pyruvate
show the reaction diagram
L-cystathionine + H2O
L-homocysteine + pyruvate + NH3
show the reaction diagram
L-cysteine
L-cysteine persulfide + pyruvate + NH4+
show the reaction diagram
-
-
-
-
?
L-cysteine + H2O
?
show the reaction diagram
L-cysteine + H2O
hydrogen sulfide + pyruvate + ?
show the reaction diagram
L-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
L-cysteine S-sulfate + H2O
?
show the reaction diagram
-
11% of the activity with L-cystine
-
?
L-cystine
L-cysteine persulfide + pyruvate + NH4+
show the reaction diagram
-
cystine is preferred over cysteine as substrate
-
-
?
L-cystine + H2O
?
show the reaction diagram
22% activity compared to L-cystathionine
-
-
?
L-cystine + H2O
thiocysteine + pyruvate + NH3
show the reaction diagram
L-djenkolate + H2O
pyruvate
show the reaction diagram
L-djenkolate + H2O
pyruvate + NH3 + ?
show the reaction diagram
most effective substrate, 154% activity compared to L-cystathionine
-
-
?
L-djenkolic acid + H2O
?
show the reaction diagram
-
18-24% of the activity with L-cystine, depending on assay method
-
?
L-homocysteine + H2O
?
show the reaction diagram
L-homolanthionine + H2O
?
show the reaction diagram
-
-
-
-
?
L-meso-lanthionine + H2O
?
show the reaction diagram
-
-
-
-
?
L-methionine
methanethiol + (2S)-2-aminobutanoic acid
show the reaction diagram
-
-
-
-
?
L-methionine + H2O
methanethiol + dimethyl disulfide + dimethyl trisulfide + 2-oxobutyrate
show the reaction diagram
L-methionine + H2O
methanethiol + NH3 + 2-oxobutanoate
show the reaction diagram
L-selenocystathionine + H2O
?
show the reaction diagram
L-serine + H2O
hydrogen sulfide + pyruvate + NH3
show the reaction diagram
MalY
-
-
?
lanthionine + H2O
L-cysteine + NH3 + pyruvate
show the reaction diagram
mixed disulfide of L-cysteine and L-homocysteine + H2O
?
show the reaction diagram
-
-
-
-
?
O-acetyl-L-serine + H2O
acetate + pyruvate + NH3
show the reaction diagram
-
-
-
-
?
O-succinyl-L-homoserine + H2O
2-oxobutyrate + succinate + NH3
show the reaction diagram
MetC
-
-
?
S-(1-hydroxyhexan-3-yl)-L-cysteine + H2O
3-mercaptohexan-1-ol + NH3 + pyruvate
show the reaction diagram
-
Str3p is able to release 0.0021 mM 3-mercaptohexan-1-ol from 2 mM concentrations of its precursor (S)-3-(hexan-1-ol)-L-cysteine
-
?
S-(2-aminoethyl)-L-Cys + H2O
?
show the reaction diagram
-
-
-
-
?
S-ethyl-L-cysteine
?
show the reaction diagram
S-ethyl-L-cysteine + H2O
?
show the reaction diagram
-
18% of the activity with L-cystine
-
?
S-ethyl-L-cysteine sulfoxide + H2O
?
show the reaction diagram
-
71% of the activity with L-cystine
-
?
S-methyl-L-Cys + H2O
?
show the reaction diagram
-
-
-
-
?
S-methyl-L-cysteine + H2O
?
show the reaction diagram
S-methylcysteine + H2O
?
show the reaction diagram
-
-
-
-
?
S-propyl-L-cysteine + H2O
?
show the reaction diagram
-
29% of the activity with L-cystine
-
?
S-[1-(2-hydroxymethyl)-1-methylbutyl]-L-cysteine + H2O
sulfanylhexanol + ?
show the reaction diagram
-
very low activity
-
-
?
S-[1-(2-hydroxymethyl)-1-methylbutyl]-L-cysteinylglycine + H2O
sulfanylhexanol + ?
show the reaction diagram
-
very low activity
-
-
?
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
cystathionine + H2O
?
show the reaction diagram
L-cystathionine + H2O
L-homocysteine + NH3 + pyruvate
show the reaction diagram
L-cystathionine + H2O
L-homocysteine + pyruvate + NH3
show the reaction diagram
L-cysteine + H2O
sulfide + NH3 + pyruvate
show the reaction diagram
A6YH85
-
-
-
?
L-methionine + H2O
methanethiol + NH3 + 2-oxobutanoate
show the reaction diagram
A6YH85
-
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
pyridoxal 5'-phosphate
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-(4-chlorophenyl)-3-[4-(6-methyl-1,3-benzothiazol-2-yl)phenyl]urea
-
reversible inhibition
1-[3-(3,4-dichlorophenyl)isoxazol-5-yl]-2-[(4-thiophen-2-ylpyrimidin-2-yl)sulfanyl]ethanone
-
reversible inhibition
2,3,5-trifluoro-N-(2-hydrazino-2-oxoethyl)-4-methoxybenzamide
-
only poor inhibitor
2-bromobenzyl (9Z)-9-(hydroxyimino)-2,7-dinitro-9H-fluorene-4-carboxylate
-
hydrolysis of the oxime to generate hydroxylamine in situ that interacted with the PLP cofactor
3,3,3,-trifluoroalanine
3,3,3-trifluoro-N-(2-methylphenyl)-2-(trifluoromethyl)propanamide
-
reversible inhibition
3,4-dichlorobenzenesulfonohydrazide
-
known to interact with the cofactor of PLP enzymes
3,5-diamino-N-(2-hydrazino-2-oxoethyl)benzamide
-
-
3-amino-N-(2-hydrazino-2-oxoethyl)naphthalene-2-carboxamide
-
only poor inhibitor
3-methyl-2-benzothiazolinone hydrazoone
-
-
4-chloro-N-[4-(6-methyl-1,3-benzothiazol-2-yl)phenyl]benzamide
-
reversible inhibition
4-methylbenzenesulfonohydrazide
-
known to interact with the cofactor of PLP enzymes
5,5'-dithiobis(2-nitrobenzoic acid)
5-aminonaphthalene-1-sulfonohydrazide
-
known to interact with the cofactor of PLP enzymes
5-chlorothiophene-2-sulfonohydrazide
-
known to interact with the cofactor of PLP enzymes
5-pyridin-2-ylthiophene-2-sulfonohydrazide
-
known to interact with the cofactor of PLP enzymes
aminoethoxyvinylglycine
ammonium sulfate
-
66% inhibition at 4 mM
aromatic disulfide
-
-
-
Beta-cyanoalanine
carboxymethoxylamine
-
irreversible
diphenyl 9-(dicyanomethylidene)-4,5-dinitro-9H-fluorene-2,7-disulfonate
-
reversible inhibition
DL-cycloserine
-
irreversible
DL-homocysteine
-
12.5 mM, 70% inhibition, competitive inhibitor
DL-Penicillamine
-
-
DL-propargylglycine
DTT
-
10 mM, 19% inhibition
glutathione
-
strong
glycine
-
73% inhibition at 4 mM
homocysteine
-
strong
hydroxylamine
iodoacetamide
iodoacetate
L-alpha-(2-aminoethoxyvinyl)glycine
L-aminoethoxyvinylglycine
-
time-dependent slow-binding, one-step mechanism
L-canaline
-
1 mM, 67% inhibition, 10 mM complete inhibition
L-cystathionine
-
substrate inhibition at concentrations above 6 mM
L-cysteine
-
12 mM, 65% inhibition after 5 min, 1 mM, 29% inhibition after 10 min, linear noncompetitive inhibitor
L-Met
-
competitive
L-methionine
-
40% inhibition at 6 mM
L-serine
-
86% inhibition at 4 mM
N-(2-hydrazino-2-oxoethyl)-2,6-dimethoxybenzamide
-
-
N-(2-hydrazino-2-oxoethyl)-2-(trifluoromethyl)benzamide
-
-
N-(2-hydrazino-2-oxoethyl)-2-naphthalen-1-ylacetamide
-
-
N-(2-hydrazino-2-oxoethyl)-2-nitrobenzamide
-
known to interact with the cofactor of PLP enzymes
N-(2-hydrazino-2-oxoethyl)-3,5-dinitrobenzamide
-
-
N-(2-hydrazino-2-oxoethyl)-3-(trifluoromethyl)benzamide
-
-
N-(2-hydrazino-2-oxoethyl)-4-(trifluoromethyl)benzamide
-
-
N-(2-hydrazino-2-oxoethyl)-4-nitrobenzamide
-
-
N-hydrazinocarbonylmethyl-2-nitrobenzamide
-
-
N-hydrazinocarbonylmethyl-2-trifluoromethylbenzamide
-
-
N-[4-(hydrazinosulfonyl)benzyl]acetamide
-
known to interact with the cofactor of PLP enzymes
NaF
-
10 mM, weak
O-acetylserine
-
strong
p-hydroxymercuribenzoate
-
1 mM, 92% inhibition
phenylhydrazine
pyridoxal
-
12.5 mM, in presence of 12 mM L-cystine 12% inhibition, in presence of 1 mM L-cystine 37% inhibition
pyruvate
-
12.5 mM, 25% inhibition
rhizobitoxine
S-adenosylmethionine
-
weak
Semicarbazide
-
-
Sodium cyanide
-
12.5 mM, 95% inhibition
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
CYS3
a bZIP transcriptional activator, is the key regulator and necessary for regulation expression of the sulfur-related genes, binds on a consensus sequence for CYS3 binding
-
NaCl
NaCl considerably enhances the reaction rate at pH 6.8; NaCl considerably enhances the reaction rate at pH 6.8
pyridoxal
-
is 23% as effective as pyridoxal 5'-phosphate
TTHA1554
-
when an 8fold excess of TTHA1554 is added to TTHA1620, the cystathionine beta-lyase activity of TTHA1620 increases approximately 2fold
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.38 - 4.2
cystathionine
1
cystine
-
estimated from steady-state measurements
5.3
D-Cys
-
-
1 - 1.4
DL-homocysteine
0.04 - 15
L-cystathionine
0.29 - 1
L-cysteine
0.0812 - 5.4
L-cystine
0.033 - 0.36
L-djenkolate
4.5
L-homolanthionine
-
-
0.83
L-meso-lanthionine
-
-
32.8 - 34.2
L-methionine
0.35
L-selenocystathionine
-
-
0.104
Lanthionine
-
-
0.47
S-(2-aminoethyl)-L-Cys
-
-
0.67
S-methyl-L-Cys
-
-
additional information
additional information
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.5
cystathionine
Salmonella enterica
-
-
0.91 - 79
L-cystathionine
1.27
L-djenkolate
Saccharomyces cerevisiae
E3VL26
in 50 mM phosphate buffer, pH 8.5, 0.02 mM pyridoxal 5'-phosphate, 1 mM EDTA, at 37C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00009 - 470
L-cystathionine
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0019 - 9.1
aminoethoxyvinylglycine
1.5
DL-homocysteine
-
-
5
L-cysteine
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.016
1-(4-chlorophenyl)-3-[4-(6-methyl-1,3-benzothiazol-2-yl)phenyl]urea
Escherichia coli
-
pH 8.5, 22C
0.00065
2,3,5-trifluoro-N-(2-hydrazino-2-oxoethyl)-4-methoxybenzamide
Escherichia coli
-
pH 8.5, 22C
0.00053
2-bromobenzyl (9Z)-9-(hydroxyimino)-2,7-dinitro-9H-fluorene-4-carboxylate
Escherichia coli
-
pH 8.5, 22C
0.00028
3,3,3-trifluoro-N-(2-methylphenyl)-2-(trifluoromethyl)propanamide
Escherichia coli
-
pH 8.5, 22C
0.0022
3,4-dichlorobenzenesulfonohydrazide
Escherichia coli
-
pH 8.5, 22C
0.082
3,5-diamino-N-(2-hydrazino-2-oxoethyl)benzamide
Escherichia coli
-
pH 8.5, 22C
0.56
3-amino-N-(2-hydrazino-2-oxoethyl)naphthalene-2-carboxamide
Escherichia coli
-
pH 8.5, 22C
0.2
4-chloro-N-[4-(6-methyl-1,3-benzothiazol-2-yl)phenyl]benzamide
Escherichia coli
-
pH 8.5, 22C
0.0018
4-methylbenzenesulfonohydrazide
Escherichia coli
-
pH 8.5, 22C
0.006
5-aminonaphthalene-1-sulfonohydrazide
Escherichia coli
-
pH 8.5, 22C
0.0034
5-pyridin-2-ylthiophene-2-sulfonohydrazide
Escherichia coli
-
pH 8.5, 22C
0.00134 - 3.5
aminoethoxyvinylglycine
0.009
diphenyl 9-(dicyanomethylidene)-4,5-dinitro-9H-fluorene-2,7-disulfonate
Escherichia coli
-
pH 8.5, 22C
0.06
N-(2-hydrazino-2-oxoethyl)-2,6-dimethoxybenzamide
Escherichia coli
-
pH 8.5, 22C
0.000079
N-(2-hydrazino-2-oxoethyl)-2-(trifluoromethyl)benzamide
Escherichia coli
-
pH 8.5, 22C
0.5
N-(2-hydrazino-2-oxoethyl)-2-naphthalen-1-ylacetamide
Escherichia coli
-
pH 8.5, 22C
0.0045
N-(2-hydrazino-2-oxoethyl)-2-nitrobenzamide
Escherichia coli
-
pH 8.5, 22C
0.0059
N-(2-hydrazino-2-oxoethyl)-3,5-dinitrobenzamide
Escherichia coli
-
pH 8.5, 22C
0.037
N-(2-hydrazino-2-oxoethyl)-3-(trifluoromethyl)benzamide
Escherichia coli
-
pH 8.5, 22C
0.015
N-(2-hydrazino-2-oxoethyl)-4-(trifluoromethyl)benzamide
Escherichia coli
-
pH 8.5, 22C
0.025
N-(2-hydrazino-2-oxoethyl)-4-nitrobenzamide
Escherichia coli
-
pH 8.5, 22C
0.034
N-[4-(hydrazinosulfonyl)benzyl]acetamide
Escherichia coli
-
pH 8.5, 22C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0091
-
-
1258
using L-cystathionine as substrate,in 50 mM phosphate buffer, pH 8.5, 0.02 mM pyridoxal 5'-phosphate, 1 mM EDTA, at 37C
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5 - 8.5
-
rapid decrease of activity above pH 8.5
8.3 - 9
-
with L-cystathionine or L-djenkolate as substrate
8.6 - 8.7
-
-
8.6
-
Tris-HCl buffer
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 6.8
MalY: 55% of maximal activity at pH 5.5, optimal activity at pH 6.0, reduced activity at pH 6.8, no activity at pH 9.0
5.5 - 9
MetC: 15% of maximal activity at pH 5.5, optimal activity at pH 9.0
7 - 10.5
using L-cystathionine as substrate, no significant activity is observed below pH 7.0 or above pH 10.5
7.2 - 9
-
pH 7.2: 36% of maximal activity, pH 8.0-9.0: optimum
7.5 - 9
-
pH 7.5: about 20% of maximal activity, pH 7.5-9.0: optimum
8.4 - 9.9
-
pH 8.4: about 55% of maximal activity, pH 9.9: about 30% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
60 - 90
-
60C: optimum, 90C: 17% of maximal activity
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.3
calculation from nucleotide sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
100000
-
gel filtration
130000
-
non-denaturing PAGE
165000
-
gel filtration
170000
195000
gel filtration
210000
-
gel filtration
280000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
-
-
homotetramer
tetramer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
-
carbohydrate content of 3%
no modification
-
enzyme is not a glycoprotein
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cocrystallization with inhibitor N-(2-hydrazino-2-oxoethyl)-2-nitrobenzamide as well as with N-(2-hydrazino-2-oxoethyl)-2,6-dimethoxybenzamide
-
sitting drop vapor diffusion method. Crystal structure of the lyase and of a complex with the reaction products of cystine cleavage at 1.8 A and 1.55 A resolution
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.8 - 9
at pH 6.8 Ctl1 still exhibits high cystathionine lyase activity, no cystathionine lyase activity is detected at pH 9.0; at pH 6.8 cystathionine lyase activity of Ctl2 is strongly reduced, no cystathionine lyase activity is detected at pH 9.0
708517
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
20 h, stable
25 - 40
-
stable, rapid decline in activity above 40C
42 - 57
Ctl1 does not lose activity after incubation at 42C for 30 min, but activity is lost at 57C; Ctl2 does not lose activity after incubation at 42C for 30 min, but activity is lost at 57C
45
-
10 min, completely stable up to
50
-
10 min, 20% loss of activity
56
30 min, inactivation; 30 min, inactivation
70
-
10 min, complete inactivation above
additional information
-
Tm values of wild-type and mutant enzymes, overview
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
addition of 15% glycerol extends the enzymatic lifetime to several days and permitted freezing of the preparation at -20C
-
inactivated by repeated freezing and thawing
-
no decrease in activity after repeated freezing, -20C, and thawing
-
not stable to lyophilization
-
rapid loss of activity during dialysis
-
repeated freezing, -20C, and thawing has no effect
-
the buffer exchange conditions to stabilize the isolated protein and maintain its enzymatic activity include 500 mM KCl and 20 mM EDTA. By including 10% (w/v) glycerol in the buffer, less than 6% of the activity is lost in a freeze-thaw cycle, compared to the 56% loss in the presence of 10 mM imidazole
the coenzyme strongly increases the protein stability. It is essential for the step involving dissociation of dimer into monomers and is not required for refolding of single monomers, but it is necessary to attain the native dimeric structure critical for the biological activities of MalY
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-10C, stable for 1 month
-
-15C or 4C, weeks to months, slow loss of activity
-
-20C or 4C, 50% glycerol, 4 months, stable
-
-20C, stable for 6 months
-
-20C, stable for months
-
-80C or at 4C, 5 days, 70% loss of activity, partially purified enzyme
-
-80C, 10 mM MOPS buffer, pH 6.5
-
-80C, 25 mM Tris-HCl, pH 7.8, 30 mM KCl, 1 mM DTT, 0.5 mM EDTA, 5% ethanediol, less than 5% loss of activity
-
-80C, stable for several months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, Toyopearl butyl-650M resin column chromatography, and DEAE column chromatography (untagged enzyme), or Ni-NTA column chromatography (His-tagged enzyme)
-
chloroplastic isoenzyme
-
cocrystallization of wild-type enzyme and mutant enzyme K223A with cystine and cysteine
-
Ni-Sepharose column chromatography
Ni2+ affinity column chromatography; Ni2+ affinity column chromatography
recombinant His-tagged enzyme from Escherichia coli strain BL21 by nickel affinity chromatography
-
recombinant His-tagged MalY from Escherichia coli by nickel affinity chromatography; recombinant His-tagged MetC from Escherichia coli by nickel affinity chromatography
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain ER1821 metC::cat by nickel affinity chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
construction of an overproducing strain of Escherichia coli
-
DNA and amino acid sequence determination and analysis, expression of His-tagged MalY in Escherichia coli; DNA and amino acid sequence determination and analysis, expression of His-tagged MetC in Escherichia coli
expressed in Escherichia coli BL21(DE3) cells; expressed in Escherichia coli BL21(DE3) cells
expressed in Saccharomyces cerevisiae strain VIN 13 and in Escherichia coli BL21(DE3) cells
expression in Escherichia coli
expression in Escherichia coli. Overexpression of the enzyme complements the methionine auxotrophy of an Escherichia coli metC mutant
-
expression of wild-type and chimeric mutant enzymes in Escherichia coli strain ER1821, in vivo complementation of methionine-auxotrophic Escherichia coli strains, lacking the genes encoding cystathionine gamma-synthase and cystathionine beta-lyase, with chimeric mutants
-
from strain MG1363 and from strain B78
gene met-2+, DNA and amino acid sequence determination and analysis, presence of CYS3 activator binding sites on the met-2+ promoter, quantitative enzyme expression analysis
gene metC or SH2636, phylogenetic tree and analysis, cloning from strains AX3 and CSC1435, expression of His-tagged enzyme in Escherichia coli strain BL21
-
mutant enzymes are expressed in Escherichia coli KS1000 cells
-
mutant enzymes are expressed in the Escherichia coli KS1000 metC:cat strain
-
overexpression in Escherichia coli
-
overexpression in Escherichia coli CAG18475
-
recloning of the gene from a transducing phage lambda carrying the structural gene for beta-cystathionase
-
recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain ER1821 metC::cat
-
the deduced amino acid sequence of beta-cystathionase shows extensive homology with that of cystathionine gamma-synthase
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
expression of met-2+ in wild-type Neurspora crassa increases 3.1fold under sulfur-limiting growth conditions as compared to the transcript levels seen under high sulfur growth conditions, i.e., repressing conditions
in a activator knockout DELTAcys-3 mutant strain, the met-2+ transcript levels are substantially reduced under either low- or high-sulfur growth conditions
STR3 transcript levels in the modified strain VIN 13 are 18fold higher at day 5 but only 50% higher by the end of grape juice fermentation. In the VIN 13 control strain, the level of STR3 transcript under the control of its native promoter increase 12.3fold between day 5 and day 15 of fermentation
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C117G
-
inactive protein, pyridoxal 5'-phosphate is not detectable in the mutant protein, enhanced susceptibility to chymotrypsin digestion
C279
-
activity is comparable with that of the native enzyme
C309A
-
inactive protein, pyridoxal 5'-phosphate is not detectable in the mutant protein, enhanced susceptibility to chymotrypsin digestion
C88A
-
activity is comparable with that of the native enzyme
D116A
-
mutant with reduced catalytic efficiency
D116N
-
mutant with reduced catalytic efficiency
F55D
-
the mutant shows 74fold reduced catalytic efficiency compared to the wild type enzyme
F55D/Y338E
-
the mutant shows 58000fold reduced catalytic efficiency compared to the wild type enzyme
K42A
-
site-directed mutagenesis, the mutant shows slightly reduced catalytic efficiency compared to the wild-type enzyme
R372A
-
mutant with reduced catalytic efficiency
R372K
-
mutant with reduced catalytic efficiency
R372L
-
mutant with reduced catalytic efficiency
R58A
-
mutant with reduced catalytic efficiency
R58K
-
mutant with reduced catalytic efficiency
R59A
-
mutant with reduced catalytic efficiency
R59K
-
mutant with reduced catalytic efficiency
S32A
-
site-directed mutagenesis, the mutant shows slightly reduced catalytic efficiency compared to the wild-type enzyme
S33A
-
site-directed mutagenesis, the mutant shows slightly reduced catalytic efficiency compared to the wild-type enzyme
W131F
-
site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme
W131F/W188F/W230F/W276F
-
site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme
W131F/W188F/W230F/W276F/W300F
-
site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme
W131F/W188F/W230F/W276F/W340F
-
site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme
W131F/W188F/W230F/W300F/W340F
-
site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme
W131F/W188F/W276F/W300F/W340F
-
site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme
W131F/W230F/W276F
-
site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme
W131F/W230F/W276F/W300F
-
site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme
W131F/W230F/W276F/W300F/W340F
-
site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme
W131F/W230F/W276F/W340F
-
site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme
W188F
-
site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by 4.7C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme
W188F/W230F/W276F/W300F/W340F
-
site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme
W230F
-
site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme
W276F
-
site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme
W300F
-
site-directed mutagenesis, below 2fold increase in KM and kcat for L-cystathionine and by less than 1.7C reduced midpoint of thermal denaturation, monitored by circular dichroism spectroscopy, compared to the wild-type enzyme
Y338E
-
the mutant shows 2850fold reduced catalytic efficiency compared to the wild type enzyme
K223A
-
inactive mutant enzyme
additional information
-
construction of 12 chimeric mutants of cystathionine gamma-synthase, EC 2.5.1.48, and cystathionine beta-lyase to probe the roles of two structurally distinct, about 25-residue segments situated in proximity to the amino and carboxy termini and located at the entrance of the active-site. The exchange of the targeted regions impairs the activity of the resulting enzymes, but does not produce a corresponding interchange of reaction specificity, catalytic efficiency of the native reactions is reduced by at least 95fold, and alpha,beta versus alpha,gamma-elimination specificity is not modified. The chimeric enzymes adopt a stable folded structure
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
urea-induced unfolding, unfolding proceeds in at least three stages.The first transition, occurring between 0 and 1 M urea, gives rise to a partially active dimeric species that binds pyridoxal 5'-phosphate. The second equilibrium transition involving dimer dissociation, release of pyridoxal 5'-phosphate and loss of lyase activity leads to the formation of a monomeric equilibrium intermediate. It is a partially unfolded molecule that retains most of the native-state secondary structure, binds significant amounts of 8-anilino-1-naphthalenesulfonic acid (a probe for exposed hydro-phobic surfaces) and tends to self-associate. The self-associated aggregates predominate at urea concentrations of 24 M for holoMalY. The third step represents the complete unfolding of the enzyme. Both holo-and apo-MalY can be successfully refolded into the active enzyme with an 85% yield. Large misfolded soluble aggregates cannot be refolded and can be responsible for the incomplete reactivation
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
the absence of the enzyme in higher organisms makes it an important target for the development of antibiotics and herbicides
food industry
medicine
nutrition
-
possible essential role of the enzyme in flavor development in cheese is suggested
Show AA Sequence (4058 entries)
Please use the Sequence Search for a certain query.