SUBN_BACNA
ID SUBN_BACNA Reviewed; 381 AA.
AC P35835;
DT 01-JUN-1994, integrated into UniProtKB/Swiss-Prot.
DT 01-JUN-1994, sequence version 1.
DT 03-AUG-2022, entry version 119.
DE RecName: Full=Subtilisin NAT;
DE EC=3.4.21.62;
DE AltName: Full=Nattokinase;
DE Flags: Precursor;
GN Name=aprN;
OS Bacillus subtilis subsp. natto.
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX NCBI_TaxID=86029;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=NC2-1;
RX PubMed=1369081; DOI=10.1271/bbb.56.1869;
RA Nakamura T., Yamagata Y., Ichishima E.;
RT "Nucleotide sequence of the subtilisin NAT gene, aprN, of Bacillus subtilis
RT (natto).";
RL Biosci. Biotechnol. Biochem. 56:1869-1871(1992).
RN [2]
RP PROTEIN SEQUENCE OF 107-381, AND CHARACTERIZATION.
RX PubMed=8280151; DOI=10.1006/bbrc.1993.2624;
RA Fujita M., Nomura K., Hong K., Ito Y., Asada A., Nishimuro S.;
RT "Purification and characterization of a strong fibrinolytic enzyme
RT (nattokinase) in the vegetable cheese natto, a popular soybean fermented
RT food in Japan.";
RL Biochem. Biophys. Res. Commun. 197:1340-1347(1993).
CC -!- FUNCTION: Subtilisin is an extracellular alkaline serine protease, it
CC catalyzes the hydrolysis of proteins and peptide amides. Subtilisin NAT
CC also has fibrinolytic activity.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=Hydrolysis of proteins with broad specificity for peptide
CC bonds, and a preference for a large uncharged residue in P1.
CC Hydrolyzes peptide amides.; EC=3.4.21.62;
CC -!- COFACTOR:
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108; Evidence={ECO:0000250};
CC Note=Binds 2 calcium ions per subunit. {ECO:0000250};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.48 mM for Suc-Ala-Ala-Pro-Phe-pNA;
CC -!- SUBUNIT: Monomer.
CC -!- SUBCELLULAR LOCATION: Secreted.
CC -!- MISCELLANEOUS: Secretion of subtilisin is associated with onset of
CC sporulation, and many mutations which block sporulation at early stages
CC affect expression levels of subtilisin. However, subtilisin is not
CC necessary for normal sporulation.
CC -!- MISCELLANEOUS: In Japan this enzyme, isolated from a process of
CC fermentation involving boiled soybeans and Bacillus natto, is used as a
CC food supplement because it is reported to reduce and prevent blood
CC clotting.
CC -!- SIMILARITY: Belongs to the peptidase S8 family. {ECO:0000305}.
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DR EMBL; D25319; BAA04989.1; -; Genomic_DNA.
DR EMBL; S51909; AAC60424.1; -; Genomic_DNA.
DR PIR; JH0778; JH0778.
DR RefSeq; WP_014479360.1; NZ_SJSU01000014.1.
DR PDB; 3VYV; X-ray; 1.36 A; A/B=107-381.
DR PDB; 4DWW; X-ray; 1.74 A; A=107-381.
DR PDB; 5GL8; X-ray; 1.80 A; A/B=107-381.
DR PDBsum; 3VYV; -.
DR PDBsum; 4DWW; -.
DR PDBsum; 5GL8; -.
DR AlphaFoldDB; P35835; -.
DR BMRB; P35835; -.
DR SMR; P35835; -.
DR MEROPS; I09.001; -.
DR MEROPS; S08.044; -.
DR PRIDE; P35835; -.
DR SABIO-RK; P35835; -.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0004252; F:serine-type endopeptidase activity; IEA:UniProtKB-EC.
DR GO; GO:0042730; P:fibrinolysis; IEA:UniProtKB-KW.
DR GO; GO:0007599; P:hemostasis; IEA:UniProtKB-KW.
DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW.
DR GO; GO:0030435; P:sporulation resulting in formation of a cellular spore; IEA:UniProtKB-KW.
DR CDD; cd07477; Peptidases_S8_Subtilisin_subset; 1.
DR Gene3D; 3.30.70.80; -; 1.
DR Gene3D; 3.40.50.200; -; 1.
DR InterPro; IPR000209; Peptidase_S8/S53_dom.
DR InterPro; IPR036852; Peptidase_S8/S53_dom_sf.
DR InterPro; IPR023827; Peptidase_S8_Asp-AS.
DR InterPro; IPR022398; Peptidase_S8_His-AS.
DR InterPro; IPR023828; Peptidase_S8_Ser-AS.
DR InterPro; IPR015500; Peptidase_S8_subtilisin-rel.
DR InterPro; IPR010259; S8pro/Inhibitor_I9.
DR InterPro; IPR037045; S8pro/Inhibitor_I9_sf.
DR InterPro; IPR034202; Subtilisin_Carlsberg-like.
DR Pfam; PF05922; Inhibitor_I9; 1.
DR Pfam; PF00082; Peptidase_S8; 1.
DR PRINTS; PR00723; SUBTILISIN.
DR SUPFAM; SSF52743; SSF52743; 1.
DR PROSITE; PS51892; SUBTILASE; 1.
DR PROSITE; PS00136; SUBTILASE_ASP; 1.
DR PROSITE; PS00137; SUBTILASE_HIS; 1.
DR PROSITE; PS00138; SUBTILASE_SER; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Blood coagulation; Calcium; Direct protein sequencing;
KW Fibrinolysis; Hemostasis; Hydrolase; Metal-binding; Protease; Secreted;
KW Serine protease; Signal; Sporulation; Zymogen.
FT SIGNAL 1..29
FT /evidence="ECO:0000250"
FT PROPEP 30..106
FT /evidence="ECO:0000269|PubMed:8280151"
FT /id="PRO_0000027175"
FT CHAIN 107..381
FT /note="Subtilisin NAT"
FT /id="PRO_0000027176"
FT DOMAIN 38..103
FT /note="Inhibitor I9"
FT /evidence="ECO:0000255"
FT DOMAIN 111..380
FT /note="Peptidase S8"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT ACT_SITE 138
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT ACT_SITE 170
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT ACT_SITE 327
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT BINDING 108
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000250"
FT BINDING 147
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000250"
FT BINDING 181
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000250"
FT BINDING 183
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000250"
FT BINDING 185
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000250"
FT BINDING 187
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000250"
FT BINDING 275
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000250"
FT BINDING 277
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000250"
FT BINDING 280
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000250"
FT HELIX 112..116
FT /evidence="ECO:0007829|PDB:3VYV"
FT HELIX 119..125
FT /evidence="ECO:0007829|PDB:3VYV"
FT STRAND 133..139
FT /evidence="ECO:0007829|PDB:3VYV"
FT STRAND 150..155
FT /evidence="ECO:0007829|PDB:3VYV"
FT STRAND 167..169
FT /evidence="ECO:0007829|PDB:3VYV"
FT HELIX 170..179
FT /evidence="ECO:0007829|PDB:3VYV"
FT STRAND 182..186
FT /evidence="ECO:0007829|PDB:3VYV"
FT STRAND 195..200
FT /evidence="ECO:0007829|PDB:3VYV"
FT HELIX 210..222
FT /evidence="ECO:0007829|PDB:3VYV"
FT STRAND 226..230
FT /evidence="ECO:0007829|PDB:3VYV"
FT HELIX 239..250
FT /evidence="ECO:0007829|PDB:3VYV"
FT STRAND 254..258
FT /evidence="ECO:0007829|PDB:3VYV"
FT TURN 274..276
FT /evidence="ECO:0007829|PDB:3VYV"
FT STRAND 280..286
FT /evidence="ECO:0007829|PDB:3VYV"
FT STRAND 304..307
FT /evidence="ECO:0007829|PDB:3VYV"
FT STRAND 309..315
FT /evidence="ECO:0007829|PDB:3VYV"
FT TURN 316..318
FT /evidence="ECO:0007829|PDB:3VYV"
FT STRAND 319..323
FT /evidence="ECO:0007829|PDB:3VYV"
FT HELIX 326..343
FT /evidence="ECO:0007829|PDB:3VYV"
FT HELIX 349..358
FT /evidence="ECO:0007829|PDB:3VYV"
FT HELIX 366..369
FT /evidence="ECO:0007829|PDB:3VYV"
FT HELIX 376..379
FT /evidence="ECO:0007829|PDB:3VYV"
SQ SEQUENCE 381 AA; 39507 MW; DAED4B16ED1BA092 CRC64;
MRSKKLWISL LFALTLIFTM AFSNMSAQAA GKSSTEKKYI VGFKQTMSAM SSAKKKDVIS
EKGGKVQKQF KYVNAAAATL DEKAVKELKK DPSVAYVEED HIAHEYAQSV PYGISQIKAP
ALHSQGYTGS NVKVAVIDSG IDSSHPDLNV RGGASFVPSE TNPYQDGSSH GTHVAGTIAA
LNNSIGVLGV APSASLYAVK VLDSTGSGQY SWIINGIEWA ISNNMDVINM SLGGPTGSTA
LKTVVDKAVS SGIVVAAAAG NEGSSGSTST VGYPAKYPST IAVGAVNSSN QRASFSSVGS
ELDVMAPGVS IQSTLPGGTY GAYNGTSMAT PHVAGAAALI LSKHPTWTNA QVRDRLESTA
TYLGNSFYYG KGLINVQAAA Q
