Information on EC 3.4.24.78 - gpr endopeptidase

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The expected taxonomic range for this enzyme is: Firmicutes

EC NUMBER
COMMENTARY
3.4.24.78
-
RECOMMENDED NAME
GeneOntology No.
gpr endopeptidase
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Endopeptidase action with P4 Glu or Asp, P1 preferably Glu > Asp, P1' hydrophobic and P2' Ala
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
germinating-specific protease, GPS
-
-
-
-
germination proteinase
-
-
-
-
germination proteinase
-
-
CAS REGISTRY NUMBER
COMMENTARY
75718-32-0
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain QN 8155
-
-
Manually annotated by BRENDA team
Bacillus megaterium QN 8155
strain QN 8155
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
GPR zymogen + H2O
?
show the reaction diagram
-
-
-
-
?
N-acetyl-Phe-Glu-Ile-Ala-Ser-Glu-Phe-Gly-Val-Asn-Leu-Gly-Pro-Asp-Ala-Thr-Ala-Arg + H2O
N-acetyl-Phe-Glu + Ile-Ala-Ser-Glu-Phe-Gly-Val-Asn-Leu-Gly-Pro-Asp-Ala-Thr-Ala-Arg
show the reaction diagram
-
-
-
?
N-acetyl-Thr-Glu-Phe-Ala + H2O
N-acetyl-Thr-Glu + Phe-Ala
show the reaction diagram
-
-
-
?
N-acetyl-Thr-Glu-Phe-Ala-Ser-Glu-Phe + H2O
N-acetyl-Thr-Glu + Phe-Ala-Ser-Glu-Phe
show the reaction diagram
-
-
-
?
small acid-soluble protein + H2O
?
show the reaction diagram
-
-
-
-
?
small acid-soluble protein + H2O
?
show the reaction diagram
-
-
-
-
?
small acid-soluble protein alpha + H2O
fragments of small acid-soluble protein alpha
show the reaction diagram
-
-
-
?
small acid-soluble protein alpha + H2O
fragments of small acid-soluble protein alpha
show the reaction diagram
-
1 cleavage site
-
-
-
small acid-soluble protein beta + H2O
fragments of small acid-soluble protein beta
show the reaction diagram
-
-
-
?
small acid-soluble protein beta + H2O
fragments of small acid-soluble protein beta
show the reaction diagram
-
1 cleavage site
-
-
-
small acid-soluble protein gamma + H2O
fragments of small acid-soluble protein gamma
show the reaction diagram
-
-
-
?
small acid-soluble protein gamma + H2O
fragments of small acid-soluble protein gamma
show the reaction diagram
-
2 or 3 cleavage sites
-
-
-
spore protein A + H2O
fragments of spore protein A
show the reaction diagram
-
specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
-
-
-
spore protein A + H2O
fragments of spore protein A
show the reaction diagram
-
specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
-
?
spore protein A + H2O
fragments of spore protein A
show the reaction diagram
Bacillus megaterium QN 8155
-
specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
-
-
-
spore protein A + H2O
fragments of spore protein A
show the reaction diagram
Bacillus megaterium QN 8155
-
specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
-
?
spore protein B + H2O
fragments of spore protein B
show the reaction diagram
-
specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
-
-
-
spore protein B + H2O
fragments of spore protein B
show the reaction diagram
-
specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
-
?
spore protein B + H2O
fragments of spore protein B
show the reaction diagram
Bacillus megaterium QN 8155
-
specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
-
-
-
spore protein B + H2O
fragments of spore protein B
show the reaction diagram
Bacillus megaterium QN 8155
-
specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
-
?
spore protein C + H2O
fragments of spore protein C
show the reaction diagram
-
specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
-
-
-
spore protein C + H2O
fragments of spore protein C
show the reaction diagram
-
specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
-
?
spore protein C + H2O
fragments of spore protein C
show the reaction diagram
Bacillus megaterium QN 8155
-
specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
small acid-soluble protein + H2O
?
show the reaction diagram
-
-
-
-
?
small acid-soluble protein alpha + H2O
fragments of small acid-soluble protein alpha
show the reaction diagram
-
1 cleavage site
-
-
-
small acid-soluble protein beta + H2O
fragments of small acid-soluble protein beta
show the reaction diagram
-
1 cleavage site
-
-
-
small acid-soluble protein gamma + H2O
fragments of small acid-soluble protein gamma
show the reaction diagram
-
2 or 3 cleavage sites
-
-
-
spore protein A + H2O
fragments of spore protein A
show the reaction diagram
Bacillus megaterium, Bacillus megaterium QN 8155
-
specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
-
-
-
spore protein B + H2O
fragments of spore protein B
show the reaction diagram
Bacillus megaterium, Bacillus megaterium QN 8155
-
specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
-
-
-
spore protein C + H2O
fragments of spore protein C
show the reaction diagram
Bacillus megaterium, Bacillus megaterium QN 8155
-
specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
-
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
P22321
gpr is likely to be regulated by metal ions
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
p-chloromercuribenzenesulfonate
-
1 mM, 39% inhibition, addition of 10 mM dithiothreitol restores activity to 100%
PMSF
-
1 mM, 68% inhibition after 1 h
HgCl2
-
1 mM, 73% inhibition, addition of 10 mM dithiothreitol restores activity to 100%
additional information
-
not inhibited by E-64, pepstatin, bestatin, EDTA, antipain, APMSF, leupeptin, aprotinin and SSI
-
additional information
-
not inhibited by EDTA, 1,10-phenanthroline, DFP, N-ethylmaleimide, iodoacetamide or by inhibitors of aspartic proteases
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.4
N-acetyl-Phe-Glu-Ile-Ala-Ser-Glu-Phe-Gly-Val-Asn-Leu-Gly-Pro-Asp-Ala-Thr-Ala-Arg
-
pH 7.4, 37°C
20
N-acetyl-Thr-Glu-Phe-Ala-Ser-Glu-Phe
-
pH 7.4, 37°C
0.014
purified small acid-soluble protein
-
-
-
20
small tetra-to heptapeptides containing a cleavage site
-
-
-
0.0014
spore protein A
-
spore protein A, Km lower than 0.014 mM
-
0.008
spore protein B
-
Km lower than 0.008 mM
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
139363 units, 1 unit is defined as a decrease in the OD600 of 0.1 per min
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
coat of dormant spores
Manually annotated by BRENDA team
Bacillus megaterium QN 8155
-
-
-
Manually annotated by BRENDA team
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
-
x * 52000 + x * 57000 + x * 60000, processed forms of preproteins, SDS-PAGE, x * 51442 + x * 55458 + x * 55585, deduced from nucleotide sequence
homotetramer
-
4 * 40000
tetramer
-
wild-type P46 nearly 100% tetrameric, P41 largely tetrameric but also with a significant percentage of lower-molecular-mass-species
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
proteolytic modification
-
gpr is synthesized in the developing spore as precursor which is termed P46, this precursor is processed, probably autocatalytically, to an active enzyme called P41 about 2 h later in sporulation
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
at 3 A resolution
-
truncated enzyme termed P30, hanging-drop vapor-diffusion, mixing 0.002 ml of protein and 0.002 ml of reservoir solution containing 32% polyethylene glycol 4000, 100 mM Tris-HCl, pH 8.3 at room temperature, crystals of P30 appear within 4 d, crystals diffract to about 3.3 A
-
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7
-
at least 2 month at 4°C in the presence of 0.1% CHAPS, no loss of activity
638930
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4
-
72 h, 80% loss of activity in the absence of CHAPS
638930
55
-
40 min at pH 7.0, no loss of activity in the presence of 0.1% CHAPS
638930
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
to homogeneity
-
Superose 12, hydroxyapatite, DEAE-5PW
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
-
subcloned into the pET23a vector, overexpression of full-length gene P46 and a truncated form encoding only P41 in Escherichia coli Bl21(DE3)pLysS
-
expression of P41 in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D127A
-
no activity of P41 against small acid-soluble spore protein, no autoprocessing of P46
D127E
-
no activity of P41 against small acid-soluble spore protein, no autoprocessing of P46
D127N
-
no activity of P41 against small acid-soluble spore protein, no autoprocessing of P46
D127S
-
no activity of P41 against small acid-soluble spore protein, no autoprocessing of P46
D153N
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
D193N
-
no activity of P41 against small acid-soluble spore protein, no autoprocessing of P46
D211N
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
D246N
-
partial activity of P41 against small acid-soluble spore protein, no autoprocessing of P46
D253N
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
D256N
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
E201Q
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
E26Q
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
E89Q
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
H125Q
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
H142Q
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
K21P
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
K223A
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
K223E
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
K223H
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
Q208E
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
R202Q
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
R224Q
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
S199A
-
activity of P41 against small acid-soluble spore protein, no autoprocessing of P46
S210A
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
S212A
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
S218A
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
S249A
-
partial activity of P41 against small acid-soluble spore protein, no autoprocessing of P46
T125N
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
T173V
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
T215V
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
T74D
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
Y72F
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
K223M
-
activity of P41 against small acid-soluble spore protein, autoprocessing of P46
additional information
-
Delta223 mutant with activity of P41 against small acid-soluble spore protein and autoprocessing of P46, Delta213-226 mutant, Delta213-227 mutant, Delta214-226 mutant and Delta214-227 mutant with no activity of P41 against small acid-soluble spore protein and no autoprocessing of P46
additional information
-
gpr-negative mutants do not rapidly proceed to vegetative growth following spore germination, as transcription is inhibited by the undegraded small acid-soluble protein
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
degradation
-
D127 and D193 are essential for activity and autoprocessing
synthesis
-
the enzyme is synthesized only during sporulation within the developing spore, and as a zymogen, ca. 2 h after its synthesis the zymogen autoprocesses to the active enzyme by intramolecular removal of 15 N-terminal residues
synthesis
-
the enzyme is synthesized only during sporulation within the developing spore, and as a zymogen, ca. 2 h after its synthesis the zymogen autoprocesses to the active enzyme by intramolecular removal of 16 N-terminal residues