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3.4.24.78
cohesin
anaphase
chromatid
securin
spindle
aneuploidy
centromeres
metaphase-to-anaphase
rad21
disjunction
kinetochore
centriole
kleisin
separase-dependent
missegregation
sister-chromatides
degradation
synthesis
The expected taxonomic range for this enzyme is: Bacteria, Archaea
Reaction Schemes
Endopeptidase action with P4 Glu or Asp, P1 preferably Glu > Asp, P1' hydrophobic and P2' Ala
Synonyms
germinating-specific protease, GPS, germination proteinase, GPR, tepA, YmfB,
more
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germinating-specific protease, GPS
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germination proteinase
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germination proteinase
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germination proteinase
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germination proteinase
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germination proteinase
Halalkalibacterium halodurans
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germination proteinase
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germination proteinase
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GPR
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GPR
Halalkalibacterium halodurans
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Endopeptidase action with P4 Glu or Asp, P1 preferably Glu > Asp, P1' hydrophobic and P2' Ala
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hydrolysis of peptide bond
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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
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?
N-acetyl-Thr-Glu-Phe-Ala + H2O
N-acetyl-Thr-Glu + Phe-Ala
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?
N-acetyl-Thr-Glu-Phe-Ala-Ser-Glu-Phe + H2O
N-acetyl-Thr-Glu + Phe-Ala-Ser-Glu-Phe
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purified small acid-soluble protein + H2O
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?
small acid-soluble protein + H2O
?
small acid-soluble protein alpha + H2O
fragments of small acid-soluble protein alpha
small acid-soluble protein beta + H2O
fragments of small acid-soluble protein beta
small acid-soluble protein gamma + H2O
fragments of small acid-soluble protein gamma
small tetra-to heptapeptides containing a cleavage site + H2O
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?
spore protein A + H2O
fragments of spore protein A
spore protein B + H2O
fragments of spore protein B
spore protein C + H2O
fragments of spore protein C
GPR zymogen + H2O
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?
GPR zymogen + H2O
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small acid-soluble protein + H2O
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small acid-soluble protein + H2O
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small acid-soluble protein + H2O
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small acid-soluble protein alpha + H2O
fragments of small acid-soluble protein alpha
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small acid-soluble protein alpha + H2O
fragments of small acid-soluble protein alpha
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1 cleavage site
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?
small acid-soluble protein beta + H2O
fragments of small acid-soluble protein beta
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small acid-soluble protein beta + H2O
fragments of small acid-soluble protein beta
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1 cleavage site
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?
small acid-soluble protein gamma + H2O
fragments of small acid-soluble protein gamma
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?
small acid-soluble protein gamma + H2O
fragments of small acid-soluble protein gamma
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2 or 3 cleavage sites
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?
spore protein A + H2O
fragments of spore protein A
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specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
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?
spore protein A + H2O
fragments of spore protein A
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specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
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?
spore protein B + H2O
fragments of spore protein B
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specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
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?
spore protein B + H2O
fragments of spore protein B
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specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
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?
spore protein C + H2O
fragments of spore protein C
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specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
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?
spore protein C + H2O
fragments of spore protein C
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specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
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?
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small acid-soluble protein + H2O
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small acid-soluble protein alpha + H2O
fragments of small acid-soluble protein alpha
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1 cleavage site
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?
small acid-soluble protein beta + H2O
fragments of small acid-soluble protein beta
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1 cleavage site
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?
small acid-soluble protein gamma + H2O
fragments of small acid-soluble protein gamma
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2 or 3 cleavage sites
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?
spore protein A + H2O
fragments of spore protein A
spore protein B + H2O
fragments of spore protein B
spore protein C + H2O
fragments of spore protein C
small acid-soluble protein + H2O
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small acid-soluble protein + H2O
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small acid-soluble protein + H2O
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spore protein A + H2O
fragments of spore protein A
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specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
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spore protein A + H2O
fragments of spore protein A
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specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
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?
spore protein B + H2O
fragments of spore protein B
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specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
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spore protein B + H2O
fragments of spore protein B
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specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
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?
spore protein C + H2O
fragments of spore protein C
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specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
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?
spore protein C + H2O
fragments of spore protein C
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specific sequence of cleavage site, R-Glu-(Phe,Ile)-Ala,Gly-Ser-Glu-R, gpr endopeptidase cleaves at the first glutamyl bond
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additional information
gpr is likely to be regulated by metal ions
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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
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not inhibited by EDTA, 1,10-phenanthroline, DFP, N-ethylmaleimide, iodoacetamide or by inhibitors of aspartic proteases
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additional information
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not inhibited by E-64, pepstatin, bestatin, EDTA, antipain, APMSF, leupeptin, aprotinin and SSI
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additional information
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not inhibited by EDTA, 1,10-phenanthroline, DFP, N-ethylmaleimide, iodoacetamide or by inhibitors of aspartic proteases
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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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additional information
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139363 units, 1 unit is defined as a decrease in the OD600 of 0.1 per min
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brenda
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brenda
Halalkalibacterium halodurans
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SwissProt
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Uniprot
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coat of dormant spores
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brenda
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brenda
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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
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51442
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x * 51442 + x * 55458 + x * 55585, deduced from nucleotide sequence
52000
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x * 52000 + x * 57000 + x * 60000, processed forms of preproteins, SDS-PAGE
55458
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x * 51442 + x * 55458 + x * 55585, deduced from nucleotide sequence
55585
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x * 51442 + x * 55458 + x * 55585, deduced from nucleotide sequence
57000
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x * 52000 + x * 57000 + x * 60000, processed forms of preproteins, SDS-PAGE
60000
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x * 52000 + x * 57000 + x * 60000, processed forms of preproteins, SDS-PAGE
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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
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x * 52000 + x * 57000 + x * 60000, processed forms of preproteins, SDS-PAGE
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x * 51442 + x * 55458 + x * 55585, deduced from nucleotide sequence
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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
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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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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
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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
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gpr-negative mutants do not rapidly proceed to vegetative growth following spore germination, as transcription is inhibited by the undegraded small acid-soluble protein
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7
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at least 2 month at 4°C in the presence of 0.1% CHAPS, no loss of activity
638930
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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
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Superose 12, hydroxyapatite, DEAE-5PW
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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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degradation
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D127 and D193 are essential for activity and autoprocessing
synthesis
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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
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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
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Dignam, S.S.; Setlow, P.
Bacillus megaterium spore protease. Action of the enzyme on peptides containing the amino acid sequence cleaved by the enzyme in vivo
J. Biol. Chem.
255
8408-8412
1980
Priestia megaterium, Priestia megaterium QN 8155
brenda
Carrillo-Martinez, Y.; Setlow, P.
Properties of Bacillus subtilis small, acid-soluble spore proteins with changes in the sequence recognized by their specific protease
J. Bacteriol.
176
5357-5363
1994
Bacillus subtilis
brenda
Ponnuraj, K.; Kelly, S.; Nessi, C.; Setlow, P.; Jedrzejas, M.J.
Crystallization and preliminary diffraction studies of a truncated form of a novel protease from spores of Bacillus megaterium
Acta Crystallogr. Sect. D
56
70-72
2000
Priestia megaterium
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Shimamoto, S.; Moriyama, R.; Sugimoto, K.; Miyata, S.; Makino, S.
Partial characterization of an enzyme fraction with protease activity which converts the spore peptidoglycan hydrolase (SleC) precursor to an active enzyme during germination of Clostridium perfringens S40 spores and analysis of a gene cluster involved in the activity
J. Bacteriol.
183
3742-3751
2001
Clostridium perfringens
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Pei, J.; Grishin, N.V.
Breaking the singleton of germination protease
Protein Sci.
11
691-697
2002
Priestia megaterium (P22321)
brenda
Setlow, P.
Endopeptidase GPR
Handbook Of Proteolytic Enzymes(Barrett,A. J. ,Rawlings,N. D. ,Woessner,J. F. ,Eds. )Academic Press
1
983-984
2004
Bacillus anthracis, Bacillus cereus, Priestia megaterium, Bacillus subtilis, Halalkalibacterium halodurans, Oceanobacillus iheyensis
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Carroll, T.M.; Setlow, P.
Site-directed mutagenesis and structural studies suggest that the germination protease, GPR, in spores of Bacillus species is an atypical aspartic acid protease
J. Bacteriol.
187
7119-7125
2005
Priestia megaterium
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Traag, B.; Pugliese, A.; Setlow, B.; Setlow, P.; Losick, R.
A conserved ClpP-like protease involved in spore outgrowth in Bacillus subtilis
Mol. Microbiol.
90
160-166
2013
Bacillus subtilis (Q99171)
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