Information on EC 1.21.4.2 - glycine reductase

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The expected taxonomic range for this enzyme is: Bacteria

EC NUMBER
COMMENTARY
1.21.4.2
-
RECOMMENDED NAME
GeneOntology No.
glycine reductase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin
show the reaction diagram
cleavage mechanism for the pyruvoyl group dependent reductase starting from cysteine; mechanism; The reaction is observed only in the direction of glycine reduction. The enzyme consists of three protein components A, B and C. Protein B contains selenocysteine and a pyruvoyl group, and is responsible for glycine binding and ammonia release. Protein A, which also contains selenocysteine, is reduced by thioredoxin, and is needed to convert the carboxymethyl group into a ketene equivalent, in turn used by protein C to produce acetyl phosphate. Only protein B distinguishes this enzyme from EC 1.21.4.3 (sarcosine reductase) and EC 1.21.4.4 (betaine reductase)
P26971
acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin
show the reaction diagram
cleavage mechanism for the pyruvoyl group dependent reductase starting from cysteine; mechanism; The reaction is observed only in the direction of glycine reduction. The enzyme consists of three protein components A, B and C. Protein B contains selenocysteine and a pyruvoyl group, and is responsible for glycine binding and ammonia release. Protein A, which also contains selenocysteine, is reduced by thioredoxin, and is needed to convert the carboxymethyl group into a ketene equivalent, in turn used by protein C to produce acetyl phosphate. Only protein B distinguishes this enzyme from EC 1.21.4.3 (sarcosine reductase) and EC 1.21.4.4 (betaine reductase)
Q9R4G7
acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin
show the reaction diagram
mechanism; The reaction is observed only in the direction of glycine reduction. The enzyme consists of three protein components A, B and C. Protein B contains selenocysteine and a pyruvoyl group, and is responsible for glycine binding and ammonia release. Protein A, which also contains selenocysteine, is reduced by thioredoxin, and is needed to convert the carboxymethyl group into a ketene equivalent, in turn used by protein C to produce acetyl phosphate. Only protein B distinguishes this enzyme from EC 1.21.4.3 (sarcosine reductase) and EC 1.21.4.4 (betaine reductase)
-
acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin
show the reaction diagram
a selenium-containing protein, selenoprotein, is essential component of the enzyme
-
acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin
show the reaction diagram
a selenium-containing protein, selenoprotein, is essential component of the enzyme
-
acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin
show the reaction diagram
the protein C component catalyses the arsenate-dependent decomposition of acetyl phosphate; the protein component C serves as the acetyl group acceptor in the overall reaction
-
acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin
show the reaction diagram
The reaction is observed only in the direction of glycine reduction. The enzyme consists of three protein components A, B and C. Protein B contains selenocysteine and a pyruvoyl group, and is responsible for glycine binding and ammonia release. Protein A, which also contains selenocysteine, is reduced by thioredoxin, and is needed to convert the carboxymethyl group into a ketene equivalent, in turn used by protein C to produce acetyl phosphate. Only protein B distinguishes this enzyme from EC 1.21.4.3 (sarcosine reductase) and EC 1.21.4.4 (betaine reductase)
-
acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin
show the reaction diagram
the protein C component catalyses the arsenate-dependent decomposition of acetyl phosphate; The reaction is observed only in the direction of glycine reduction. The enzyme consists of three protein components A, B and C. Protein B contains selenocysteine and a pyruvoyl group, and is responsible for glycine binding and ammonia release. Protein A, which also contains selenocysteine, is reduced by thioredoxin, and is needed to convert the carboxymethyl group into a ketene equivalent, in turn used by protein C to produce acetyl phosphate. Only protein B distinguishes this enzyme from EC 1.21.4.3 (sarcosine reductase) and EC 1.21.4.4 (betaine reductase)
-
acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin
show the reaction diagram
The reaction is observed only in the direction of glycine reduction. The enzyme consists of three protein components A, B and C. Protein B contains selenocysteine and a pyruvoyl group, and is responsible for glycine binding and ammonia release. Protein A, which also contains selenocysteine, is reduced by thioredoxin, and is needed to convert the carboxymethyl group into a ketene equivalent, in turn used by protein C to produce acetyl phosphate. Only protein B distinguishes this enzyme from EC 1.21.4.3 (sarcosine reductase) and EC 1.21.4.4 (betaine reductase)
-
acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin
show the reaction diagram
The reaction is observed only in the direction of glycine reduction. The enzyme consists of three protein components A, B and C. Protein B contains selenocysteine and a pyruvoyl group, and is responsible for glycine binding and ammonia release. Protein A, which also contains selenocysteine, is reduced by thioredoxin, and is needed to convert the carboxymethyl group into a ketene equivalent, in turn used by protein C to produce acetyl phosphate. Only protein B distinguishes this enzyme from EC 1.21.4.3 (sarcosine reductase) and EC 1.21.4.4 (betaine reductase); the thioredoxin system is involved in the electron transport from reduced pyridine nucleotides to protein A, i.e. in the electron flow between protein of glycine decarboxylase and glycine reductase complex
-
acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin
show the reaction diagram
the 48000 Da subunit of protein component C catalyses the arsenate-dependent decomposition of actetyl phosphate, a possible role of the 57000 Da subunit of protein component C could be the involvement in the reductive dehydration which leads to the cleavage of the protein A-bound carboxymethyl-selenoether to ketene and oxidized protein A; the protein C component catalyses the arsenate-dependent decomposition of acetyl phosphate; The reaction is observed only in the direction of glycine reduction. The enzyme consists of three protein components A, B and C. Protein B contains selenocysteine and a pyruvoyl group, and is responsible for glycine binding and ammonia release. Protein A, which also contains selenocysteine, is reduced by thioredoxin, and is needed to convert the carboxymethyl group into a ketene equivalent, in turn used by protein C to produce acetyl phosphate. Only protein B distinguishes this enzyme from EC 1.21.4.3 (sarcosine reductase) and EC 1.21.4.4 (betaine reductase)
-
acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin
show the reaction diagram
Cys 359 of GrdD is the thiol responsible for the formation of the acetyl thioester during catalysis of arsenate-dependent hydrolysis of acetyl phosphate; GrdD of protein component C catalyses the arsenate-dependent decomposition of acetyl phosphate, whereas GrdC completely inactive; The reaction is observed only in the direction of glycine reduction. The enzyme consists of three protein components A, B and C. Protein B contains selenocysteine and a pyruvoyl group, and is responsible for glycine binding and ammonia release. Protein A, which also contains selenocysteine, is reduced by thioredoxin, and is needed to convert the carboxymethyl group into a ketene equivalent, in turn used by protein C to produce acetyl phosphate. Only protein B distinguishes this enzyme from EC 1.21.4.3 (sarcosine reductase) and EC 1.21.4.4 (betaine reductase)
-
acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin
show the reaction diagram
glycine pathway is used for acetate synthesis
-
acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin
show the reaction diagram
mechanism, thiols are present in protein C that is acetylated during reaction; The reaction is observed only in the direction of glycine reduction. The enzyme consists of three protein components A, B and C. Protein B contains selenocysteine and a pyruvoyl group, and is responsible for glycine binding and ammonia release. Protein A, which also contains selenocysteine, is reduced by thioredoxin, and is needed to convert the carboxymethyl group into a ketene equivalent, in turn used by protein C to produce acetyl phosphate. Only protein B distinguishes this enzyme from EC 1.21.4.3 (sarcosine reductase) and EC 1.21.4.4 (betaine reductase)
-
acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin
show the reaction diagram
cleavage mechanism for the pyruvoyl group dependent reductase starting from cysteine; mechanism; The reaction is observed only in the direction of glycine reduction. The enzyme consists of three protein components A, B and C. Protein B contains selenocysteine and a pyruvoyl group, and is responsible for glycine binding and ammonia release. Protein A, which also contains selenocysteine, is reduced by thioredoxin, and is needed to convert the carboxymethyl group into a ketene equivalent, in turn used by protein C to produce acetyl phosphate. Only protein B distinguishes this enzyme from EC 1.21.4.3 (sarcosine reductase) and EC 1.21.4.4 (betaine reductase)
Clostridium sticklandii HF
-
-
acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin
show the reaction diagram
glycine pathway is used for acetate synthesis
Eubacterium acidaminophilum al-2
-
-
acetyl phosphate + NH3 + thioredoxin disulfide + H2O = glycine + phosphate + thioredoxin
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
purine nucleobases degradation I (anaerobic)
-
SYSTEMATIC NAME
IUBMB Comments
acetyl-phosphate ammonia:thioredoxin disulfide oxidoreductase (glycine-forming)
The reaction is observed only in the direction of glycine reduction. The enzyme from Eubacterium acidaminophilum consists of subunits A, B and C. Subunit B contains selenocysteine and a pyruvoyl group, and is responsible for glycine binding and ammonia release. Subunit A, which also contains selenocysteine, is reduced by thioredoxin, and is needed to convert the carboxymethyl group into a ketene equivalent, in turn used by subunit C to produce acetyl phosphate. Only subunit B distinguishes this enzyme from EC 1.21.4.3 (sarcosine reductase) and EC 1.21.4.4 (betaine reductase).
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
glycine reductase
-
-
CAS REGISTRY NUMBER
COMMENTARY
39307-24-9
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
in Brachyspira pilosicoli glycine reductase is part of a glycine reductase complex system consisting of cluster of nine genes
-
-
Manually annotated by BRENDA team
Clostridium difficile
expression induced by presence of glycine. Growth depends on ATP generation due to coupled fermentation of corresponding Stickland amino acid pairs in otherwise limiting basal medium. Enzyme and D-proline reductase are the key selenoenzymes induced by presence of Stickland amino acid pairs
-
-
Manually annotated by BRENDA team
strain HF, DSM 519T from DSMZ
SwissProt
Manually annotated by BRENDA team
three protein system consisting of proteins A,B, and C
-
-
Manually annotated by BRENDA team
Clostridium sticklandii HF
strain HF, DSM 519T from DSMZ
SwissProt
Manually annotated by BRENDA team
; protein B complex of glycine reductase has peroxidase activity using substrate dithiothreitol and hydroperoxide
-
-
Manually annotated by BRENDA team
DSM 5388T, growth on serine
SwissProt
Manually annotated by BRENDA team
three protein system consisting of proteins A,B, and C
-
-
Manually annotated by BRENDA team
Eubacterium acidaminophilum al-2
strain al-2
-
-
Manually annotated by BRENDA team
three protein system consisting of proteins A,B, and C
-
-
Manually annotated by BRENDA team
strain CDK
-
-
Manually annotated by BRENDA team
three protein system consisting of proteins A,B, and C
-
-
Manually annotated by BRENDA team
Treponema denticola CDK
strain CDK
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
-
-
-
-
-
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
-
-
-
-
?
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
-
-
-
-
-, ?
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
-
-
-
-
-
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
-
-
-
-
?
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
-
-
-
-
-
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
-
-
-
-
?
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
Q9R4G7
-
-
-
-, ?
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
-
-
-
-
-
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
-
-
-
-
?
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
P26971
-
-
-
-, ?
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
-
enzyme activity is NADPH-dependent but not dithioerythritol-dependent
-
-
?
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
-
reduction of glycine action is coupled with formation of ATP from ADP
-
?
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
-
reduction of glycine action is coupled with formation of ATP from ADP
-
-
?
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
Eubacterium acidaminophilum al-2
-
-
-
-
-, ?
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
Treponema denticola CDK
-
-
-
-
-, ?
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
Clostridium sticklandii HF
P26971
-
-
-
-, ?
dithiothreitol + cumene hydroperoxide
?
show the reaction diagram
-
peroxidase activity of enzyme
-
-
?
glycine + phosphate + thioredoxin
acetyl phosphate + ammonia + thioredoxin disulfide
show the reaction diagram
-
-
-
-
?
glycine + phosphate + thioredoxin
acetyl phosphate + ammonia + thioredoxin disulfide
show the reaction diagram
-
conservation of energy as acetyl phosphate
-
-
?
additional information
?
-
-
Se-carboxymethyl selenprotein A is a substrate of protein C
-
-
-
additional information
?
-
-
Se-carboxymethyl selenprotein A is a substrate of protein C
-
-
-
additional information
?
-
-
the B protein complex, consisting of the selenocysteine-containing GrdB subunit and two subunits, which are derived from the GrdE proprotein, shows 1.7 U/mg peroxidase activity with DTT and cumene hydroperoxide as substrates, the protein exhibits DTT- as well as NADPH-dependent peroxidase activity, overview
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
-
-
-
-
-
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
-
-
-
-
-
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
-
-
-
-
-
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
-
-
-
-
-
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
Q9R4G7
-
-
-
-
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
-
-
-
-
-
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
P26971
-
-
-
-
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
Eubacterium acidaminophilum al-2
-
-
-
-
-
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
Treponema denticola CDK
-
-
-
-
-
acetyl phosphate + ammonia + thioredoxin disulfide
glycine + phosphate + thioredoxin
show the reaction diagram
Clostridium sticklandii HF
P26971
-
-
-
-
glycine + phosphate + thioredoxin
acetyl phosphate + ammonia + thioredoxin disulfide
show the reaction diagram
-
conservation of energy as acetyl phosphate
-
-
?
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Mg2+
-
required for decomposition of acetyl phosphate by protein C
Mg2+
-
not absolutely required, but increases activity by about 20%, the highest enzyme activity in presence of 10 mM MgCl2
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
(NH4)2SO4
-
inhibits the ability of protein component C to catalyse the arsenate-dependent decomposition of acetyl phosphate
(NH4)2SO4
-
70% loss of activity at 300 mM; inhibits the ability of protein component C to catalyse the arsenate-dependent decomposition of acetyl phosphate
acetyl phosphate
-
inhibits protein C activity, although a substrate
Bromoacetate
-
75Se-labeled protein A preparation is inactivated at pH 6, 25°C, for 10 min in presence of 10 mM bromoacetate by about 25%
iodoacetate
-
the ability of protein component C to catalyse the arsenate-dependent decomposition of acetyl phosphate is inhibited, but protein C is protected from inactivation by treatment acetyl phosphate
iodoacetate
-
the ability of protein component C to catalyse the arsenate-dependent decomposition of acetyl phosphate is inhibited, but protein C is protected from inactivation by treatment acetyl phosphate
KBH4
-
inactivates protein B, very little effect on fraction C
KCl
-
the ability of protein component C to catalyse the arsenate-dependent decomposition of acetyl phosphate is inhibited
NH2OH
-
inhibits protein B and fraction C
additional information
-
no effect: CaCl2, CoCl2, MnCl2
-
additional information
-
inactivation of selenoprotein A from Clostridium purinolyticum by sheep antibodies elicited to selenoprotein A from Clostridium stricklandii
-
additional information
-
antibodies raised against the thioredoxin reductase-like flavoprotein or thioredoxin inhibit to a high extent
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
arsenate
-
absolutely required for protein C activity
arsenate
-
absolutely required for protein C activity
thiols
-
such as dithiothreitol required for decomposition of acetyl phosphate by protein C
thiols
-
not absolutely required, but increases activity by about 20%, the highest enzyme activity in presence of 10 mM dithioerythritol
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
additional information
-
additional information
-
kinetics of the DTT-dependent peroxidase activity of the protein B complex
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.5
-
-
protein B of enzyme
51.3
-
-
protein C of enzyme
352
-
Q9R4G7
substrate-specific selenoprotein B of enzyme
additional information
-
-
-
additional information
-
-
the protein B complex shows 1.7 U/mg peroxidase activity with DTT and cumene hydroperoxide as substrates
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.5
-
-
of the arsenate dependent decomposition of acetyl phosphate, Tris/HCl buffer
7.5
-
-
peroxidase activity assay at
10
-
-
of the arsenate dependent decomposition of acetyl phosphate, piperazin buffer
additional information
-
-
pI of protein component C: 5.7
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
22
-
peroxidase activity assay at
40
-
-
of the arsenate dependent decomposition of acetyl phosphate
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
evidence for presence of a clostridial-type enzyme
Manually annotated by BRENDA team
Treponema denticola CDK
-
evidence for presence of a clostridial-type enzyme
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
420000
-
-
protein component C, gel filtration
additional information
-
-
molecular mass is depending on the salt concentration present
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
dimer
P26971
selenoprotein B, alpha,beta, 1 * 21162 + 1 * 26295, mass spectroscopy, amino acid sequence
dimer
Clostridium sticklandii HF
-
selenoprotein B, alpha,beta, 1 * 21162 + 1 * 26295, mass spectroscopy, amino acid sequence
-
hexamer
Q9R4G7
selenoprotein B, alpha,beta,gamma, 2 * 22000 + 2 * 25000 + 2 * 47000
monomer
-
selenoprotein A, 1* 25000 Da, but the two selenoproteins differ in amino acid sequence
monomer
-
selenoprotein A, 1 * 17022, mass spectroscopy; selenoprotein A, 1* 25000 Da, but the two selenoproteins differ in amino acid sequence
monomer
-
1 * 17011 calculated from amino acid sequence; selenoprotein A, 1 * 17022, mass spectroscopy
monomer
-
selenoprotein A, 1 * 18500, SDS-PAGE, the two selenoproteins exhibit very similar N-terminal amino acid sequences
monomer
-
selenoprotein A, 1 * 16700, amino acid sequence; selenoprotein B, 1 * 47000, amino acid sequence
monomer
-
selenoprotein A, 1 * 18000; selenoprotein B, 1 * 45000, SDS-PAGE
multimer
-
protein C, x * 40000 + x * 54000
octamer
-
protein C, alpha,beta, 4 * 57000 + 4 * 48000, SDS-PAGE
monomer
Treponema denticola CDK
-
selenoprotein A, 1 * 18000; selenoprotein B, 1 * 45000, SDS-PAGE
-
additional information
-
three protein system consisting of protein A (17000 Da), protein B (47000 Da or 48000 Da, later processed into two proteins of 22000 and 25000 Da), and protein C (40000 or 54000 Da)
additional information
-
heterologous enzyme is protected from degradation by full-length GrdE or by GrdE domains; the enzyme consists of three subunits A, B, and C. The protein B-complex consists of the selenocysteine-containing GrdB subunit, subunit B, and two subunits, which derive from the GrdE proprotein, one of which shows peroxidase activity and protects the sensitive selenoproteins in the organism
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
glycoprotein
-
selenoprotein A is a glycoprotein
no glycoprotein
-
selenoprotein A component of enzyme is not a glycoprotein
glycoprotein
-
selenoprotein A is a glycoprotein
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
47
-
-
30% loss of activity of protein C after heating for 10 min, at pH 7.0, in the presence of EDTA, with and without Mg2+
62
64
-
protein C inactivated
68
-
-
80% loss of activity of protein C after heating for 10 min, at pH 7.0, in the presence of EDTA, with and without Mg2+
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
the ability of protein component C to catalyse the arsenate-dependent decomposition of acetyl phosphate is inhibited by alkylation selenoprotein A alkylated at pH 6 with bromoacetate is active as a component of the enzyme complex
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20°C, no significant loss of activity during storage
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
of selenoprotein component PA by 75Se incorporation
-
of protein C of enzyme
-
of pyruvoyl group-forming protein grdE
P26971
of selenoprotein A of enzyme
-
; copurification of recombinant GrdE with recombinant Strep-tagged GrdB, native DTT-dependent peroxidase activity 14fold by anion exchange and hydrophobic interaction chromatography, ammonium sulfate fractionation, and gel filtration
-
of protein C of enzyme
-
of protein C of enzyme, recombinant enzyme
-
of selenoprotein B of enzyme
Q9R4G7
of selenoprotein component PA by 75Se incorporation
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
cloning of selenocystein-containing protein A: grdA and substrate-specific selenoprotein B: grdB
-
of the pyruvoyl group-forming proproteins grdE, expression in Escherichia coli
P26971
cloning and sequencing of a new gene region, encoding a proprotein for the beta and alpha subunits of selenoprotein B: grdE, selenoprotein A: grdA and selenium-containing gamma subunit of selenoprotein B: grdB
Q9R4G7
cloning of two subunits of protein C: grdC1 and grdD1, expression in Escherichia coli
-
expression of wild-type and mutant B protein complex components in Escherichia coli strain XL1-Blue
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C242A
P26971
no cleavage on the N-terminal side of a cysteine
C242S
P26971
cleavage on the N-terminal side of a cysteine under similar conditions with more extended half-times than wild-type enzyme
C242T
P26971
cleavage on the N-terminal side of a cysteine under similar conditions with more extended half-times than wild-type enzyme
C242A
Clostridium sticklandii HF
-
no cleavage on the N-terminal side of a cysteine
-
C242S
Clostridium sticklandii HF
-
cleavage on the N-terminal side of a cysteine under similar conditions with more extended half-times than wild-type enzyme
-
C242T
Clostridium sticklandii HF
-
cleavage on the N-terminal side of a cysteine under similar conditions with more extended half-times than wild-type enzyme
-
C353
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mutation of potentially redox-active motif UxxCxxC, 44% of wild-type peroxidase activity
C356
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mutation of potentially redox-active motif UxxCxxC, 40% of wild-type peroxidase activity
C359A
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grdD of protein component C, mutant enzyme completely inactive, accessible to iodoacetate only under native conditions, suggesting that Cys359 of GrdD is the thiol responsible for the formation of the acetyl thioester during catalysis of arsenate-dependent hydrolysis of acetyl phosphate
U350
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mutation of potentially redox-active motif UxxCxxC, 60% of wild-type peroxidase activity
C98S
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grdD of protein component C, activity is unchanged, accessible to iodoacetate only after denaturation
additional information
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mutation of potentially redox-active motif UxxCxxC results in still significant, but decreased peroxidase activity; mutation of the potentially redox-active UxxCxxC motif in subunit GrdB of the B protein complex results in still signifiant, but decreased peroxidase activity, overview
Renatured/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
denaturation with SDS and 2-mercaptoethanol for 15 min at 100°C does not lead to degradation of protein PA
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