AORSN_ASPOR
ID AORSN_ASPOR Reviewed; 652 AA.
AC Q8NK92; Q2UFT5;
DT 01-MAR-2005, integrated into UniProtKB/Swiss-Prot.
DT 02-MAY-2006, sequence version 2.
DT 03-AUG-2022, entry version 100.
DE RecName: Full=Aorsin;
DE EC=3.4.21.-;
DE Flags: Precursor;
GN Name=aorO {ECO:0000312|EMBL:BAB97387.1}; ORFNames=AO090026000083;
OS Aspergillus oryzae (strain ATCC 42149 / RIB 40) (Yellow koji mold).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Eurotiomycetes;
OC Eurotiomycetidae; Eurotiales; Aspergillaceae; Aspergillus;
OC Aspergillus subgen. Circumdati.
OX NCBI_TaxID=510516;
RN [1] {ECO:0000305, ECO:0000312|EMBL:BAB97387.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 216-244; 516-530 AND
RP 604-612, FUNCTION, COFACTOR, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP PROPERTIES, SUBCELLULAR LOCATION, AND MUTAGENESIS OF GLU-301; ASP-305;
RP ASP-426; SER-569 AND ASP-610.
RC STRAIN=ATCC 42149 / RIB 40;
RX PubMed=12519073; DOI=10.1042/bj20021691;
RA Lee B.R., Furukawa M., Yamashita K., Kanasugi Y., Kawabata C., Hirano K.,
RA Ando K., Ichishima E.;
RT "Aorsin, a novel serine proteinase with trypsin-like specificity at acidic
RT pH.";
RL Biochem. J. 371:541-548(2003).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 42149 / RIB 40;
RX PubMed=16372010; DOI=10.1038/nature04300;
RA Machida M., Asai K., Sano M., Tanaka T., Kumagai T., Terai G., Kusumoto K.,
RA Arima T., Akita O., Kashiwagi Y., Abe K., Gomi K., Horiuchi H.,
RA Kitamoto K., Kobayashi T., Takeuchi M., Denning D.W., Galagan J.E.,
RA Nierman W.C., Yu J., Archer D.B., Bennett J.W., Bhatnagar D.,
RA Cleveland T.E., Fedorova N.D., Gotoh O., Horikawa H., Hosoyama A.,
RA Ichinomiya M., Igarashi R., Iwashita K., Juvvadi P.R., Kato M., Kato Y.,
RA Kin T., Kokubun A., Maeda H., Maeyama N., Maruyama J., Nagasaki H.,
RA Nakajima T., Oda K., Okada K., Paulsen I., Sakamoto K., Sawano T.,
RA Takahashi M., Takase K., Terabayashi Y., Wortman J.R., Yamada O.,
RA Yamagata Y., Anazawa H., Hata Y., Koide Y., Komori T., Koyama Y.,
RA Minetoki T., Suharnan S., Tanaka A., Isono K., Kuhara S., Ogasawara N.,
RA Kikuchi H.;
RT "Genome sequencing and analysis of Aspergillus oryzae.";
RL Nature 438:1157-1161(2005).
CC -!- FUNCTION: Serine endopeptidase which hydrolyzes a range of fluorogenic
CC peptide substrates containing the basic residues arginine or lysine at
CC the P1 position and prefers paired basic resides. Also hydrolyzes
CC clupeine and salmine, activates plasminogen and converts trypsinogen to
CC trypsin. {ECO:0000269|PubMed:12519073}.
CC -!- COFACTOR:
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC Evidence={ECO:0000269|PubMed:12519073};
CC Note=Binds 1 Ca(2+) ion per subunit. {ECO:0000269|PubMed:12519073};
CC -!- ACTIVITY REGULATION: Inhibited by antipain and leupeptin.
CC {ECO:0000269|PubMed:12519073}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.7 uM for Boc-Gln-Arg-Arg-MCA {ECO:0000269|PubMed:12519073};
CC KM=3.9 uM for Boc-Leu-Arg-Arg-MCA {ECO:0000269|PubMed:12519073};
CC KM=2.8 uM for Boc-Gly-Arg-Arg-MCA {ECO:0000269|PubMed:12519073};
CC KM=10.0 uM for Boc-Leu-Lys-Arg-MCA {ECO:0000269|PubMed:12519073};
CC KM=26.8 uM for Boc-Gly-Lys-Arg-MCA {ECO:0000269|PubMed:12519073};
CC KM=58.0 uM for Boc-Leu-Thr-Arg-MCA {ECO:0000269|PubMed:12519073};
CC KM=84.5 uM for Boc-Leu-Gly-Arg-MCA {ECO:0000269|PubMed:12519073};
CC KM=88.3 uM for Boc-Gln-Gly-Arg-MCA {ECO:0000269|PubMed:12519073};
CC Vmax=12.8 nmol/sec/mg enzyme with Z-Arg-Arg-MCA as substrate
CC {ECO:0000269|PubMed:12519073};
CC Note=The highest catalytic efficiency is observed for Boc-Leu-Lys-
CC Arg-MCA. {ECO:0000269|PubMed:12519073};
CC pH dependence:
CC Optimum pH is 4.0. Inactive below pH 3.0 and above pH 6.5. The half-
CC life (t1/2) values for activity loss at 30 degrees Celsius are 150
CC min at pH 6.6, 5.5 min at pH 6.8 and 14 min at pH 2.4.
CC {ECO:0000269|PubMed:12519073};
CC Temperature dependence:
CC Remains fully active after heating at 50 degrees Celsius and pH 4.0
CC for 10 min. Retains 65% of its activity after heating at 55 degrees
CC Celsius for 10 min. The half-life value for loss of activity at 60
CC degrees Celsius and pH 4.0 is 3.5 min. {ECO:0000269|PubMed:12519073};
CC -!- SUBCELLULAR LOCATION: Secreted, extracellular space
CC {ECO:0000269|PubMed:12519073}.
CC -!- DEVELOPMENTAL STAGE: Activity found in solid culture only. Increases at
CC the initial growth stage and decreases in the late growth stage.
CC {ECO:0000269|PubMed:12519073}.
CC -!- PTM: N-glycosylated. {ECO:0000269|PubMed:12519073}.
CC -!- PTM: O-glycosylated. {ECO:0000303|PubMed:12519073, ECO:0000305}.
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DR EMBL; AB084899; BAB97387.1; -; Genomic_DNA.
DR EMBL; AP007159; BAE59580.1; -; Genomic_DNA.
DR RefSeq; XP_001821582.1; XM_001821530.2.
DR AlphaFoldDB; Q8NK92; -.
DR SMR; Q8NK92; -.
DR MEROPS; S53.007; -.
DR EnsemblFungi; BAE59580; BAE59580; AO090026000083.
DR GeneID; 5993610; -.
DR KEGG; aor:AO090026000083; -.
DR VEuPathDB; FungiDB:AO090026000083; -.
DR HOGENOM; CLU_013783_4_0_1; -.
DR OMA; DFELAYP; -.
DR Proteomes; UP000006564; Chromosome 3.
DR GO; GO:0005576; C:extracellular region; NAS:UniProtKB.
DR GO; GO:0005615; C:extracellular space; IEA:UniProtKB-SubCell.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0004252; F:serine-type endopeptidase activity; IMP:UniProtKB.
DR GO; GO:0006508; P:proteolysis; IDA:UniProtKB.
DR CDD; cd04056; Peptidases_S53; 1.
DR CDD; cd11377; Pro-peptidase_S53; 1.
DR Gene3D; 3.40.50.200; -; 1.
DR InterPro; IPR036852; Peptidase_S8/S53_dom_sf.
DR InterPro; IPR015366; S53_propep.
DR InterPro; IPR030400; Sedolisin_dom.
DR Pfam; PF09286; Pro-kuma_activ; 1.
DR SMART; SM00944; Pro-kuma_activ; 1.
DR SUPFAM; SSF52743; SSF52743; 1.
DR PROSITE; PS51695; SEDOLISIN; 1.
PE 1: Evidence at protein level;
KW Calcium; Direct protein sequencing; Glycoprotein; Hydrolase; Metal-binding;
KW Protease; Reference proteome; Secreted; Serine protease; Signal; Zymogen.
FT SIGNAL 1..22
FT /evidence="ECO:0000255"
FT PROPEP 23..215
FT /note="Removed in mature form"
FT /evidence="ECO:0000255, ECO:0000269|PubMed:12519073"
FT /id="PRO_0000027368"
FT CHAIN 216..652
FT /note="Aorsin"
FT /evidence="ECO:0000269|PubMed:12519073"
FT /id="PRO_0000027369"
FT DOMAIN 225..651
FT /note="Peptidase S53"
FT REGION 177..211
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 301
FT /note="Charge relay system"
FT /evidence="ECO:0000250|UniProtKB:P42790"
FT ACT_SITE 305
FT /note="Charge relay system"
FT /evidence="ECO:0000250|UniProtKB:P42790"
FT ACT_SITE 569
FT /note="Charge relay system"
FT /evidence="ECO:0000250|UniProtKB:P42790"
FT BINDING 610
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000269|PubMed:12519073"
FT BINDING 611
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000250"
FT BINDING 629
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000250"
FT BINDING 631
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /evidence="ECO:0000250"
FT CARBOHYD 112
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT CARBOHYD 218
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT CARBOHYD 247
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT CARBOHYD 331
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT CARBOHYD 445
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT CARBOHYD 613
FT /note="N-linked (GlcNAc...) asparagine"
FT /evidence="ECO:0000255"
FT MUTAGEN 301
FT /note="E->D: 1000-fold decrease in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:12519073"
FT MUTAGEN 301
FT /note="E->Q: 7000-fold decrease in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:12519073"
FT MUTAGEN 305
FT /note="D->E: 2000-fold decrease in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:12519073"
FT MUTAGEN 305
FT /note="D->N: 20-fold decrease in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:12519073"
FT MUTAGEN 426
FT /note="D->E: 40000-fold decrease in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:12519073"
FT MUTAGEN 426
FT /note="D->N: 8000-fold decrease in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:12519073"
FT MUTAGEN 569
FT /note="S->T: 20000-fold decrease in catalytic efficiency."
FT /evidence="ECO:0000269|PubMed:12519073"
FT MUTAGEN 610
FT /note="D->E,N: Active. Loss of calcium binding."
FT /evidence="ECO:0000269|PubMed:12519073"
FT CONFLICT 118
FT /note="Q -> R (in Ref. 1; BAB97387)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 652 AA; 70501 MW; 865BB1A1BC8235EE CRC64;
MRPLSHLSFF NGLLLGLSAL SAATSVVHER REATSSNWVK RARVNPSDKH VVRIGLTQSS
LEEAHDLLMD VSNPSSPNYA RFYSADEVAA KFAPSTETVN EVQNWLTEKG INASRVAQTQ
NHGWLVFHAT SKEIENLFDT TYYEYHNRKT GKKAIACEQY HVPASVQKHI DYVHPGVNLN
PSSGKPSSIR RRAAASKKTK LPARGPRPIQ QHDVKGLNVT NCDQLITPEC IRALYKIPSA
RAAPHPNNSL GIFEEGDYYA QEDLDLFFKT FAKDIPQGTH PIPAFIDGAE APVPVTKAGG
ESDLDFELAY PIVHPQSITL YQTDDANWAS NTTGFLNTFL DALDGSYCTY CAYGECGNDP
SLDPVYPDDA GYDGQLMCGV FKPTNVISVS YGEQENDLPA NYQQRQCMEF LKLGLQGVSV
LFASGDNGVA GPPGDGNSVN GCLNNGTVFS PAFPNSCPYI TNVGATKVYP GYTVSQPESA
VYDPDGLYSY ASGGGFSNIY PIPDYQAEAV ATYFKDHNPP YPYYEGAENL GKNGGLYNRL
GRGYPDVAAN GDNIAVFNGG EFGSSGGTSA STPIFASIIN RIIDERLAVG KGPVGFINPV
LYKNPSVLND ITNGTNPGCG TDGFSTAPGW DPATGLGTPN YPKMLKLWLD LP
