Information on EC 4.2.1.22 - cystathionine beta-synthase

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

EC NUMBER
COMMENTARY hide
4.2.1.22
-
RECOMMENDED NAME
GeneOntology No.
cystathionine beta-synthase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-serine + L-homocysteine = L-cystathionine + H2O
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C-S bond formation
-
-
-
-
elimination
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Biosynthesis of antibiotics
-
-
Cysteine and methionine metabolism
-
-
cysteine metabolism
-
-
Glycine, serine and threonine metabolism
-
-
L-cysteine biosynthesis III (from L-homocysteine)
-
-
L-homocysteine and L-cysteine interconversion
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
L-serine hydro-lyase (adding homocysteine; L-cystathionine-forming)
A pyridoxal-phosphate protein. A multifunctional enzyme: catalyses beta-replacement reactions between L-serine, L-cysteine, cysteine thioethers, or some other beta-substituted alpha-L-amino acids, and a variety of mercaptans.
CAS REGISTRY NUMBER
COMMENTARY hide
9023-99-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
cystathionine beta-synthase
UniProt
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
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-
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Manually annotated by BRENDA team
vervet monkey
-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
stump-tailed macaque
-
-
Manually annotated by BRENDA team
long-tailed macaque
-
-
Manually annotated by BRENDA team
japanese macaque
-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
crested gibbon
-
-
Manually annotated by BRENDA team
Northern white-cheeked gibbon
-
-
Manually annotated by BRENDA team
Bonobo-pygmy chimpanzee
-
-
Manually annotated by BRENDA team
common chimpanzee
-
-
Manually annotated by BRENDA team
Hamadryas baboon
-
-
Manually annotated by BRENDA team
orang utan
-
-
Manually annotated by BRENDA team
Steinernema bibionis
-
-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
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-
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
-
cystathionine beta-synthase belongs to the fold II family of pyridoxal 5'-phosphate enzymes
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
beta-Chloro-DL-alanine + DL-homocysteine
?
show the reaction diagram
-
-
-
-
-
beta-Cyano-DL-alanine + DL-homocysteine
?
show the reaction diagram
-
-
-
-
-
cysteine + ?
H2S + ?
show the reaction diagram
DL-Serine-O-sulfate + DL-homocysteine
?
show the reaction diagram
-
-
-
-
-
L-allothreonine + homocysteine
? + H2O
show the reaction diagram
-
-
-
?
L-Cysteine + 1-butanethiol
?
show the reaction diagram
L-Cysteine + 1-mercapto-2-propanol
HOOC-CH(NH2)-CH2-S-CH2-CH(OH)-CH3 + H2S
show the reaction diagram
L-Cysteine + 1-pentanethiol
?
show the reaction diagram
L-Cysteine + 2-mercaptoethanol
HOOC-CH(NH2)-CH2-S-CH2-CH2OH + H2S
show the reaction diagram
L-cysteine + 2-mercaptoethanol
S-hydroxyethyl-L-cysteine + H2S
show the reaction diagram
-
-
-
?
L-Cysteine + cysteamine
?
show the reaction diagram
L-Cysteine + dithioerythritol
?
show the reaction diagram
L-Cysteine + dithiothreitol
?
show the reaction diagram
L-Cysteine + DL-homocysteine
Cystathionine + H2S
show the reaction diagram
L-cysteine + L-homocysteine
L-cystathionine + H2S
show the reaction diagram
L-Cysteine + monothioglycerol
?
show the reaction diagram
L-serine + cysteamine
L-thialysine
show the reaction diagram
-
-
-
-
?
L-serine + H2O
NH3 + pyruvate
show the reaction diagram
-
wild-type enzyme shows no beta-elimination reaction, beta elimination is only detectable in the following mutants: T81A, S82A, T85A, Q157A, Q157E, Q157H, Y158F
-
?
L-Serine + homocysteine
?
show the reaction diagram
L-Serine + homocysteine
Cystathionine + H2O
show the reaction diagram
L-Serine + HS-
Cysteine + OH-
show the reaction diagram
L-serine + L-cysteine
?
show the reaction diagram
-
-
-
-
?
L-serine + L-homocysteine
?
show the reaction diagram
-
-
-
-
r
L-serine + L-homocysteine
cystathionine + H2O
show the reaction diagram
L-serine + L-homocysteine
L-cystathionine + H2O
show the reaction diagram
O-acetyl-L-serine + L-homocysteine
?
show the reaction diagram
-
poor substrate for both the wild type and the N- and C-terminally truncated enzyme 71400 CBS
-
-
?
S-Methyl-L-cysteine + DL-homocysteine
?
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
L-Serine + homocysteine
?
show the reaction diagram
L-serine + L-homocysteine
cystathionine + H2O
show the reaction diagram
L-serine + L-homocysteine
L-cystathionine + H2O
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
pyridoxal 5'-phosphate
S-adenosyl-L-methionine
additional information
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
contains 0.38 Ca atoms/subunit
Co2+
-
97.1% Fe2+ and 2.9% Co2+ in wild-type FeCBS enzyme, 8% Fe2+ and 92% Co2+ in the CoCBS enzyme variant
Co3+
-
cobalt-substituted variant of hCBS, i.e. Co hCBS, in which CoPPIX replaces FePPIX, i.e. heme. Co(III) hCBS is a unique Co-substituted heme protein: the Co(III) ion is 6-coordinate, low-spin, diamagnetic, and bears a cysteine(thiolate) as one of its axial ligands. Electronic absorption and MCD spectra of the Co-substituted heme protein, overview. Co(III) hCBS is slowly reduced to Co(II) hCBS, which contains a 5-coordinate, low-spin, S = 1/2 Co-porphyrin that does not retain the cysteine(thiolate) ligand. This form of Co(II)hCBS binds NO but not CO. Co(II) hCBS is reoxidized in the air to form a new Co(III) form, which does not contain a cysteine(thiolate) ligand. Maintaining the native heme ligation motif of wild-type Fe hCBS (Cys/His) is essential in maintaining maximal activity in Co hCBS
Fe3+
-
-
Zn2+
-
contains 0.35 Zn atoms/subunit
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,4-Dichlorophenol
alizarin
Amino-oxyacetate
-
complete inhibition at 0.05 mM
aminooxyacetic acid
beta-chloro-L-alanine
-
-
Bithionol
carbon monoxide
closantel
Co3+
-
Co3+ has 30-60% of the specific activity of Fe3+-CBS
cyanide
-
-
Dichlorophene
Dithionite
-
2fold decrease in enzyme activity due to altered oxidation state of the heme
ellagic acid
Hexachlorophene
Hg2+
-
reactivity of Co(III) hCBS with HgCl2 is consistent with a loss of the cysteine(thiolate) ligand. 2-Mercaptoethanol is unable to reverse the Hg-induced ligand switch, in contrast to some other heme-thiolate proteins
hydroxylamine
iodoacetate
-
-
L-cystathione
-
product inhibition
L-homocysteine
-
substrate inhibition
Mn3+
-
Mn3+ has 30-60% of the specific activity of Fe3+-CBS
nitric oxide
p-chloromercuribenzoate
-
-
peroxynitrite
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exposure to peroxynitrite does not modify bound pyridoxal 5'-phosphate but leads to nitration of Trp208, Trp43 and Tyr223 and alterations in the heme environment including loss of thiolate coordination, conversion to high-spin and bleaching, with no detectable formation of oxoferryl compounds nor promotion of one-electron processes
Purpurogallin
quercetin
Rafoxanide
regulatory domain
-
exerts an inhibitory effect on the enzyme, deletion is correlated with a 1fold increase in catalytic activity
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titanium citrate
-
2fold decrease in enzyme activity due to altered oxidation state of the heme
-
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
AdoMet
-
allosteric regulator, 1mol per mole of monomeric subunit activates the enzyme 2fold
betaine
-
activates, effects on wild-type and mutant enzymes, overview
delta-aminolevulinic acid
-
activates, effects on wild-type and mutant enzymes, overview
Ethionine
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i.e. 2-amino-4-(ethylthio)butyric acid, an methionine analogue, which is converted to S-adenosyl-ethionine in vivo and activates CBS, treatment increased liver CBS activity 4.0fold in wild-type mice
glycerol
-
activates, effects on wild-type and mutant enzymes, overview
heme
-
regulatory role
pyridoxal 5'-phosphate
S-adenosyl-L-methionine
S-adenosylhomocysteine
-
stimulates activity 1.1fold at 0.48 mM
sinefungin
-
stimulates activity 1.28fold at 1 mM
sodium nitroprusside
-
enhances activity by interacting with cysteine residues, N-ethylmaleimide abolishes this effect
tumor necrosis factor-alpha
-
leads to cleavage of the enzyme to a truncated form and therefore increases the activity, 50% increase of activity after treatment of HepG2 cells for 16 h
-
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.72 - 24
2-mercaptoethanol
5.6 - 6.6
cysteamine
68
DL-homocysteine
-
cosubstrate L-Ser
0.05 - 67
homocysteine
0.13 - 36
L-Cys
0.083 - 0.9
L-cystathionine
2 - 6.11
L-cysteine
0.16 - 5
L-homocysteine
0.91 - 8
L-Ser
0.7 - 27.1
L-serine
additional information
additional information
-
pre-steady-state kinetic analysis of enzyme-monitored turnover during cystathionine beta-synthase-catalyzed H2S generation, overview
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5.9 - 34
homocysteine
0.0045 - 6.08
L-cystathionine
0.04 - 4.39
L-cysteine
0.024 - 21.5
L-homocysteine
7.97
L-Ser
Saccharomyces cerevisiae
-
-
0.052 - 45
L-serine
additional information
additional information
Saccharomyces cerevisiae
-
the kcat for the generation of H2S by cystathionine beta-synthase of 55/s at 37C, via the condensation of cysteine and homocysteine is 18fold faster than that for the beta-elimination and rehydration to form serine of 3/s-
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.005 - 7.5
L-cystathionine
6.3 - 80
L-homocysteine
0.031 - 25
L-serine
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5.6
CO
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37C under anaerobic conditions
2.3
cyanide
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37C
2.1
L-homocysteine
-
recombinant 6-His-tagged enzyme, in 50 mM Tris (pH 8.6), at 25C
0.32
NO
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.15
peroxynitrite
Homo sapiens
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-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.045
-
recombinant 6-His-tagged enzyme from crude extract, in 50 mM Tris (pH 8.6), at 25C
0.68
-
without pyridoxal 5'-phosphate, without S-adenosyl-L-methionine, mutant R266M
1.3
-
full-length ferrous mutant enzyme R224A, at 37C and pH 8; full-length ferrous mutant enzyme R51A, at 37C and pH 8
1.91
-
without pyridoxal 5'-phosphate, without S-adenosyl-L-methionine, mutant R266K
2.2
-
full-length ferric mutant enzyme R224A, at 37C and pH 8; full-length ferric mutant enzyme R51A, at 37C and pH 8
2.3
-
wild-type enzyme
2.31
-
without pyridoxal 5'-phosphate, without S-adenosyl-L-methionine, mutant H67A
2.6
-
-
2.65
-
wild-type enzyme in the presence of S-adenosyl-L-methionine
2.8
-
full-length ferrous wild type enzyme, at 37C and pH 8
3.17
-
recombinant 6-His-tagged enzyme after purification, in 50 mM Tris (pH 8.6), at 25C
3.5
-
-
3.6
-
without pyridoxal 5'-phosphate, without S-adenosyl-L-methionine, wild-type
3.8
-
truncated ferrous mutant enzyme R224A, at 37C and pH 8
3.9
-
with pyridoxal 5'-phosphate, with S-adenosyl-L-methionine, mutant R266M
3.95
-
D144N mutant in the presence of S-adenosyl-L-methionine
4
-
truncated ferric mutant enzyme R51A, at 37C and pH 8
4.6
-
recombinant enzyme
5.6
-
-
5.71
-
with pyridoxal 5'-phosphate, with S-adenosyl-L-methionine, mutant R266K
6
-
truncated ferric mutant enzyme R224A, at 37C and pH 8
8
-
truncated ferrous wild type enzyme, at 37C and pH 8
10.3
-
full-length ferric wild type enzyme, at 37C and pH 8
105.2
-
purified recombinant CoCBS, pH 8.6, 37C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4
-
cystathionine beta-synthase assay at
7.9 - 8.3
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enzyme form alpha
8.4 - 9
-
-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 10.5
-
-
6.5 - 10
-
-
6.5 - 9.5
-
activity increases with increasing pH, C52S and H65R mutant
6.5 - 10
-
-
7.2 - 9.3
-
pH 7.2: about 45% of maximal activity, pH 9.3: about 75% of maximal activity, full-length Fe(III)-enzyme and truncated Fe(III) CBS-45
7.3 - 9.5
-
pH 7.3: about 45% of maximal activity, pH 9.5: about 80% of maximal activity, full-length and the C-terminally truncated enzyme (CBS 1-413)
8.3 - 9.5
-
pH 8.3: about 45% of maximal activity, pH 9.5: about 85% of maximal activity N- and C-terminally truncated enzyme (BS 71-400)
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
wild-type CBS is therminally activated at 55C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
cystathionine beta-synthase is associated with the generation and/or differentiation of the radial glia/astrocyte lineage cells in the developing central nervous system
Manually annotated by BRENDA team
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cystathionine beta-synthase is associated with the generation and/or differentiation of the radial glia/astrocyte lineage cells in the developing central nervous system
Manually annotated by BRENDA team
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-
Manually annotated by BRENDA team
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highest cystathionine beta-synthase protein presence in cornea, conjunctiva and iris, followed by retina and optic nerve. Cystathionine beta-synthase may be a oxidative defense enzyme in the eye tissue, in particular in the segments of the eye where constant environmental oxidative stress is imposed
Manually annotated by BRENDA team
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-
Manually annotated by BRENDA team
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ubiquitously expressed in the ovary with the strongest expression in follicular cells at all stages. In late antral follicles, cystathionine beta-synthase expression is markedly higher in granulosa cells located close to the antrum and in cumulus cells around the oocyte
Manually annotated by BRENDA team
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cystathionine beta-synthase activities in wild-type individuals, and in hetero-, and homozygote cystathionine beta-synthase mutants, overview
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
sumoylated CBS is present in the nucleus where it is associated with the nuclear scaffold
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
36800
-
gel filtration
38000
-
gel filtration
50000 - 130000
-
existence in multiple molecular forms of MW 50000-130000, 235000, 500000, gel filtration
62800
-
gel filtration
63000
-
His-tagged enzyme, SDS-PAGE
70000
-
gel filtration, truncated enzyme, dimer
94000
-
gel filtration
100000
-
aged enzyme extract, gel filtration
115000
-
aged enzyme extract, gel filtration
119000
-
fresh enzyme extract, gel filtration
235000
250000
288000
-
fresh and aged enzyme extract, gel filtration
290000
-
fresh enzyme extract, gel filtration
400000
-
gel filtration, truncated enzyme, octamer
486000
-
octamer, gel filtration
500000
-
existence in multiple molecular forms of MW 50000-130000, 235000, 500000, gel filtration
1330000
-
multimer, gel filtration
additional information
-
high tendency for aggregation
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
-
truncated CBS, X-ray crystallography
homotetramer
multimer
octamer
oligomer
-
full length subunits for tetramers and higher oligomers
tetramer
trimer
-
3 * 20200, calculated from sequence
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
sumoylation
-
CBS is modified by the small ubiquitin-like modifier-1 protein (SUMO-I) under both in vitro and in vivo conditions. Deletion analysis of CBS indicates that the C-terminal regulatory domain is required for interaction with proteins in the sumoylation machinery. Sumoylated CBS is present in the nucleus where it is associated with the nuclear scaffold
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapour diffusion method, 100 mM phosphate-acetate pH 4.2, 200 mM NaCl and 20% (w/v) PEG 8000
-
enzyme free or in complex with a carbanion and an aminoacrylate intermediate, X-ray diffraction structure determination and analysis at 1.70 A and 1.55 A resolution, respectively
catalytic core, hanging drop vapor diffusion method
-
mutant lacking the S-adenosyl-L-methionine binding site and heme-free crystals, hanging drop vapor diffusion method
-
truncated form
-
with heme and pyridoxal 5'-phosphate, hanging drop vapor diffusion method
-
crystal structures of both mercury- and iron-bound TA0289 (1.52.0 A resolution) reveals a dimeric protein whose intersubunit contacts are formed exclusively by the alpha-helices of two cystathionine beta-synthase subdomains, whereas the C-terminal domain has a classical Zn ribbon planar architecture
-
nanodropletvapor diffusion method, crystal structure of a tandem cystathionine-beta-synthase domain protein (TM0935) from Thermotoga maritima at 1.87 A resolution
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
48
-
stable up to
55
-
melting point of wild type and S466L mutant
additional information
-
dissociation of tetrameric enzyme to dimeric form, initiated by proteolysis decreases thermostability
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
2-mercaptoethanol stabilizes
binding of S-adenosyl-L-methionine stabilizes cystathionine beta-synthase against degradation. Under pathological conditions with reduced levels of S-adenosyl-L-methionine (human hepatocellular carcinoma), level of cystathionine beta-synthase is diminished. This decrease in cystathionine beta-synthase level correlates with reduced glutathione that is, in turn, associated with increased vulnerability to oxidative stress. Posttranslational regulation of cystathionine beta-synthase stability by S-adenosyl-L-methionine provides a mechanism for achieving coordinate changes in cellular methylation and antioxidant status that is observed in a number of disease states
-
binding of S-adenosyl-L-methionine stabilizes cystathionine beta-synthase against degradation. Under pathological conditions with reduced levels of S-adenosyl-L-methionine (mouse model for chronic steatohepatitis), level of cystathionine beta-synthase is diminished. This decrease in cystathionine beta-synthase level correlates with reduced glutathione that is, in turn, associated with increased vulnerability to oxidative stress. Posttranslational regulation of cystathionine beta-synthase stability by S-adenosyl-L-methionine provides a mechanism for achieving coordinate changes in cellular methylation and antioxidant status that is observed in a number of disease states
-
high protein concentration stabilizes
inactivation by aging
-
lyophilization inactivates
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-15C, 40% glycerol, 5 weeks stable
-
-20C, 0.02 M potassium phosphate buffer, pH 7.8, 1 mM EDTA, 1 month stable
-
-20C, 0.1 M Tris-HCl buffer, pH 7.5, 0.1 mM pyridoxal 5'-phosphate
-
-20C, potassium phosphate buffer, pH 7.8, EDTA
-
-80C, potassium phosphate buffer, pH 6.5, 1 mM 2-mercaptoethanol, 3 years, no loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
CBSdeltaC143
-
full length enzyme exhibits a strong tendency toward aggregation during the course of purification as well as in the purified state
-
full length protein and truncated variant lacking 143 amino acid residues at the C-terminus
-
glutathione-linked resin chromatography, HiTrap Q Fast Flow column chromatography, GST-Sepharose column chromatography, and Superose 12 gel-filtration
-
glutathione-Sepharose column chromatography
-
GST Trap FF column chromatography and Mono Q column chromatography
-
N-terminal cystathionine beta-synthase domain fused to the C-terminal Zn ribbon domain overexpressed in Escherichia coli
Ni-NTA column chromatography
-
recombinant cystathionine beta-synthase from Escherichia coli strain BL21(DE3) by anion exchange and hydroxyapatite chromatography
-
recombinant GST-tagged wild-type and Co-subsituted CBS from Escherichia coli strain Rosetta 2 (DE3) by glutathione affinity and anion exchange chromatography
-
recombinant mutant 45CBS and wild-type CBS from Escherichia coli to homogeneity
-
truncated human CBS lacking 143 amino acids at the C-terminus is purified as a fusion protein with glutathione S-transferase using the Escherichia coli expression vector pGEXCBSN. The protein is purified through affinity chromatography with glutathione sepharose and anion exchange chromatography. The glutathione Stransferase tag was cleaved by limited proteolysis using thrombin
-
two patient-derived forms S466L and I435T, 95% purity
-
using a glutathione sepharose column
-
using Ni-NTA chromatography
-
wild type and truncated human cystathionine beta-synthase expressed in Escherichia coli
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a mutant form of the human cystathionine beta-synthase protein, I278T, is expressed in Saccharomyces cerevisiae
-
a truncated form of CBS comprising the catalytic core is used for mutational analysis
-
CBS gene encoding the truncated yCBS (residues 1-353), lacking the regulatory domain, is expressed with a C-terminal, 6-His affinity tag in Escherichia coli
-
CDCP2 is expressed in Escherichia coli BL21 (DE3) cells, selenomethionine-substituted CDCP2 is expressed using B834 (DE3) cells
-
eight CBS mutants are expressed in Escherichia coli in the presence of chemical chaperones such as ethanol, dimethyl sulfoxide, or trimethylamine-N-oxide
-
expressed as a GST-fusion protein in Escherichia coli
-
expressed in Escherichia coli
-
expressed in Escherichia coli BL21(DE3) cells
-
expressed in Escherichia coli strain DH10B
-
expressed in heme-deficient strains of Saccharomyces cerevisiae and Escherichia coli
-
expression in CHO/duk- cell line
-
expression in Escherichia coli
-
expression in yeast or in Escherichia coli
-
expression of cystathionine beta-synthase in Escherichia coli strain BL21(DE3)
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expression of human mutant CBS proteins in Saccharomyces cerevisiae reveals that the disease causing mutation severely inhibits enzyme activity and cannot support growth of yeast on cysteine-free media. The osmolyte chemical chaperones glycerol, trimethylamine-N-oxide, dimethylsulfoxide, proline or sorbitol, when added to yeast media, allows growth on cysteine-free media and causes increased enzyme activity from I278T mutant protein. The increase in enzyme activity is associated with stabilization of the tetramer form of the enzyme. This effect is not specific to yeast, as addition of the chaperone glycerol results in increased I278T activity when the enzyme is produced either in Escherichia coli or in a coupled in vitro transcription/translation reaction. No stimulation of specific activity is observed when chaperones are added directly to purified I278T indicating that the presence of chemical chaperones is required during translation
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expression of mutant 45CBS and wild-type CBS in Escherichia coli
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expression of wild type and truncated human cystathionine beta-synthase enzyme in Escherichia coli
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expression of wild-type and Co-subsituted CBS as GST-tagged enzymes in Escherichia coli strain Rosetta 2 (DE3)
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full-length CBS and truncated enzyme containing residues 1-397 are expressed in Escherichia coli BL21 cells
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fusion protein with glutathione S-transferase
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genotyping of wild-type and mutant CBSs, determination of mutations in patients with homocystinuria due to CBS deficiency, overview
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in a recombinant expression system (pGEX4T1/hCBSDELTAC143) that produces a fusion protein with glutathione S-transferase
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mutant enzymes K102N, P78R and P78R/K102N are expressed in Escherichia coli and purified as glutathione S-transferase fusion proteins using recombinant expression
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N-terminal cystathionine beta-synthase domain fused to the C-terminal Zn ribbon domain is overexpressed in Escherichia coli
the truncated human CBS enzyme consisting of residue 1-397 is expressed in the glutathione S-transferase fusion expression system
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transgenic mice that contain the human cystathionine beta-synthase cDNA under control of the zinc-inducible metallothionein promoter (Tg-CBS). In the presence of zinc, Tg-CBS mice have a 2fold to 4fold increase in liver and kidney cystathionine beta-synthase activity compared with nontransgenic littermates
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truncated form fused with glutathione S-transferase
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truncated human CBS lacking 143 amino acids at the C-terminus is expressed as a fusion protein with glutathione S-transferase in the Escherichia coli
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truncated protein lacking the C-terminal domain is expressed
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wild type enzyme is expressed in Mus musculus and mutant enzyme S466L is expressed in Saccharomyces cerevisiae strain WY35 and in Mus musculus
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
butyrate significantly increases CBS protein expression, butyrate-stimulated CBS expression is not changed by trichostatin A
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decreased expression after exogenous addition alltrans-retinoic acid
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expression level of CBS in 120 hepatocellular carcinoma specimens evaluated by RT-PCR is markedly lower than in surrounding non-cancerous liver. Reduced CBS expression is significantly correlated with high tumor stage. A survival analysis shows that a significantly shorter overall survival time is observed in patients with reduced CBS expression
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in hemizygous (+/-) CBS knockout mice the remaining CBS monoallele is up-regulated in mice when fed a taurine-deficient diet to produce additional CBS mRNA
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mice at the unusual age of 28 months even have a higher hippocampal enzyme expression than young mice
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protein level and activity increase with incubation time, upon stimulation, and similar to intracellular homocysteine, depending on intra- and extracellular homocysteine and glutathione concentrations
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the enzyme is overexpressed in primary epithelial ovarian cancer and ovarian cancer cell lines
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A331V
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mutation effects can be suppressed in a yeast assay by the deletion of the regulatory domain of the protein
C15S
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mutagenesis does not affect catalysis or S-adenosyl-L-methionine activation but significantly reduces aggregation of the purified enzyme in vitro
C272A
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2fold lower heme content, 2fold lower specific activity, 2fold higher activity in the presence of S-adenosyl-L-methionine
C274S
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2fold lower heme content, 2fold lower specific activity, 2fold higher activity in the presence of S-adenosyl-L-methionine
C431S
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mutagenesis results in a constitutively activated form of CBS that can not be further activated by either S-adenosyl-L-methionine or thermal activation
C52A
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reduced heme content, pyridoxal phosphate content comparable to wild-type enzyme, low catalytic activity
C52S
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reduced heme content, pyridoxal phosphate content comparable to wild-type enzyme, low catalytic activity