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 hide
3.4.24.78
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RECOMMENDED NAME
GeneOntology No.
gpr endopeptidase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
Endopeptidase action with P4 Glu or Asp, P1 preferably Glu > Asp, P1' hydrophobic and P2' Ala
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
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CAS REGISTRY NUMBER
COMMENTARY hide
75718-32-0
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
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Manually annotated by BRENDA team
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Manually annotated by BRENDA team
strain QN 8155
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
spores from a ymfB/gpr double mutant are more delayed in spore outgrowth and more impaired in small acid-soluble protein degradation than were spores from a gpr single mutant
physiological function
YmfB is an orphan ClpP protease involved in spore outgrowth that is dedicated to the degradation of a specialized family of small protein substrates. The function of YmfB is masked by the germination protease Gpr
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
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
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-
-
?
N-acetyl-Thr-Glu-Phe-Ala + H2O
N-acetyl-Thr-Glu + Phe-Ala
show the reaction diagram
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-
-
?
N-acetyl-Thr-Glu-Phe-Ala-Ser-Glu-Phe + H2O
N-acetyl-Thr-Glu + Phe-Ala-Ser-Glu-Phe
show the reaction diagram
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?
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 beta + H2O
fragments of small acid-soluble protein beta
show the reaction diagram
small acid-soluble protein gamma + H2O
fragments of small acid-soluble protein gamma
show the reaction diagram
spore protein A + H2O
fragments of spore protein A
show the reaction diagram
spore protein B + H2O
fragments of spore protein B
show the reaction diagram
spore protein C + H2O
fragments of spore protein C
show the reaction diagram
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
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
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1 cleavage site
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small acid-soluble protein beta + H2O
fragments of small acid-soluble protein beta
show the reaction diagram
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1 cleavage site
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small acid-soluble protein gamma + H2O
fragments of small acid-soluble protein gamma
show the reaction diagram
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2 or 3 cleavage sites
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spore protein A + H2O
fragments of spore protein A
show the reaction diagram
spore protein B + H2O
fragments of spore protein B
show the reaction diagram
spore protein C + H2O
fragments of spore protein C
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
gpr is likely to be regulated by metal ions
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
HgCl2
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1 mM, 73% inhibition, addition of 10 mM dithiothreitol restores activity to 100%
p-chloromercuribenzenesulfonate
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1 mM, 39% inhibition, addition of 10 mM dithiothreitol restores activity to 100%
PMSF
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1 mM, 68% inhibition after 1 h
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.4
N-acetyl-Phe-Glu-Ile-Ala-Ser-Glu-Phe-Gly-Val-Asn-Leu-Gly-Pro-Asp-Ala-Thr-Ala-Arg
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pH 7.4, 37°C
20
N-acetyl-Thr-Glu-Phe-Ala-Ser-Glu-Phe
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pH 7.4, 37°C
0.014
purified small acid-soluble protein
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20
small tetra-to heptapeptides containing a cleavage site
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0.0014
spore protein A
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spore protein A, Km lower than 0.014 mM
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0.008
spore protein B
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Km lower than 0.008 mM
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
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139363 units, 1 unit is defined as a decrease in the OD600 of 0.1 per min
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
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coat of dormant spores
Manually annotated by BRENDA team
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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x * 51442 + x * 55458 + x * 55585, deduced from nucleotide sequence; x * 52000 + x * 57000 + x * 60000, processed forms of preproteins, SDS-PAGE
homotetramer
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4 * 40000
tetramer
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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 hide
LITERATURE
proteolytic modification
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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
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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
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pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
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at least 2 month at 4°C in the presence of 0.1% CHAPS, no loss of activity
638930
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
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72 h, 80% loss of activity in the absence of CHAPS
55
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40 min at pH 7.0, no loss of activity in the presence of 0.1% CHAPS
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
Superose 12, hydroxyapatite, DEAE-5PW
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to homogeneity
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
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expression of P41 in Escherichia coli
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subcloned into the pET23a vector, overexpression of full-length gene P46 and a truncated form encoding only P41 in Escherichia coli Bl21(DE3)pLysS
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D127A
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no activity of P41 against small acid-soluble spore protein, no autoprocessing of P46
D127E
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no activity of P41 against small acid-soluble spore protein, no autoprocessing of P46
D127N
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no activity of P41 against small acid-soluble spore protein, no autoprocessing of P46
D127S
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no activity of P41 against small acid-soluble spore protein, no autoprocessing of P46
D153N
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
D193N
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no activity of P41 against small acid-soluble spore protein, no autoprocessing of P46
D211N
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
D246N
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partial activity of P41 against small acid-soluble spore protein, no autoprocessing of P46
D253N
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
D256N
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
E201Q
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
E26Q
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
E89Q
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
H125Q
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
H142Q
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
K21P
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
K223A
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
K223E
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
K223H
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
K223M
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
Q208E
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
R202Q
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
R224Q
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
S199A
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activity of P41 against small acid-soluble spore protein, no autoprocessing of P46
S210A
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
S212A
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
S218A
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
S249A
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partial activity of P41 against small acid-soluble spore protein, no autoprocessing of P46
T125N
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
T173V
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
T215V
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
T74D
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
Y72F
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activity of P41 against small acid-soluble spore protein, autoprocessing of P46
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
degradation
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D127 and D193 are essential for activity and autoprocessing
synthesis
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