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Protein

Subtilisin NAT

Gene

aprN

Organism
Bacillus subtilis subsp. natto
Status
Reviewed-Annotation score: Annotation score: 4 out of 5-Experimental evidence at protein leveli

Functioni

Subtilisin is an extracellular alkaline serine protease, it catalyzes the hydrolysis of proteins and peptide amides. Subtilisin NAT also has fibrinolytic activity.

Catalytic activityi

Hydrolysis of proteins with broad specificity for peptide bonds, and a preference for a large uncharged residue in P1. Hydrolyzes peptide amides.

Cofactori

Ca2+By similarityNote: Binds 2 calcium ions per subunit.By similarity

Kineticsi

  1. KM=0.48 mM for Suc-Ala-Ala-Pro-Phe-pNA

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Metal bindingi108 – 1081Calcium 1By similarity
    Active sitei138 – 1381Charge relay systemBy similarity
    Metal bindingi147 – 1471Calcium 1By similarity
    Active sitei170 – 1701Charge relay systemBy similarity
    Metal bindingi181 – 1811Calcium 1; via carbonyl oxygenBy similarity
    Metal bindingi183 – 1831Calcium 1By similarity
    Metal bindingi185 – 1851Calcium 1; via carbonyl oxygenBy similarity
    Metal bindingi187 – 1871Calcium 1; via carbonyl oxygenBy similarity
    Metal bindingi275 – 2751Calcium 2; via carbonyl oxygenBy similarity
    Metal bindingi277 – 2771Calcium 2; via carbonyl oxygenBy similarity
    Metal bindingi280 – 2801Calcium 2; via carbonyl oxygenBy similarity
    Active sitei327 – 3271Charge relay systemBy similarity

    GO - Molecular functioni

    GO - Biological processi

    Complete GO annotation...

    Keywords - Molecular functioni

    Hydrolase, Protease, Serine protease

    Keywords - Biological processi

    Blood coagulation, Fibrinolysis, Hemostasis, Sporulation

    Keywords - Ligandi

    Calcium, Metal-binding

    Enzyme and pathway databases

    SABIO-RKP35835.

    Protein family/group databases

    MEROPSiS08.044.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    Subtilisin NAT (EC:3.4.21.62)
    Alternative name(s):
    Nattokinase
    Gene namesi
    Name:aprN
    OrganismiBacillus subtilis subsp. natto
    Taxonomic identifieri86029 [NCBI]
    Taxonomic lineageiBacteriaFirmicutesBacilliBacillalesBacillaceaeBacillus

    Subcellular locationi

    GO - Cellular componenti

    Complete GO annotation...

    Keywords - Cellular componenti

    Secreted

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Signal peptidei1 – 2929By similarityAdd
    BLAST
    Propeptidei30 – 106771 PublicationPRO_0000027175Add
    BLAST
    Chaini107 – 381275Subtilisin NATPRO_0000027176Add
    BLAST

    Keywords - PTMi

    Zymogen

    Interactioni

    Subunit structurei

    Monomer.

    Structurei

    Secondary structure

    1
    381
    Legend: HelixTurnBeta strand
    Show more details
    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Helixi112 – 1165Combined sources
    Helixi119 – 1257Combined sources
    Beta strandi133 – 1397Combined sources
    Beta strandi150 – 1556Combined sources
    Beta strandi167 – 1693Combined sources
    Helixi170 – 17910Combined sources
    Beta strandi182 – 1865Combined sources
    Beta strandi195 – 2006Combined sources
    Helixi210 – 22213Combined sources
    Beta strandi226 – 2305Combined sources
    Helixi239 – 25012Combined sources
    Beta strandi254 – 2585Combined sources
    Turni274 – 2763Combined sources
    Beta strandi280 – 2867Combined sources
    Beta strandi304 – 3074Combined sources
    Beta strandi309 – 3157Combined sources
    Turni316 – 3183Combined sources
    Beta strandi319 – 3235Combined sources
    Helixi326 – 34318Combined sources
    Helixi349 – 35810Combined sources
    Helixi366 – 3694Combined sources
    Helixi376 – 3794Combined sources

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    3VYVX-ray1.36A/B107-381[»]
    4DWWX-ray1.74A107-381[»]
    ProteinModelPortaliP35835.
    SMRiP35835. Positions 36-381.
    ModBaseiSearch...
    MobiDBiSearch...

    Family & Domainsi

    Domains and Repeats

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Domaini133 – 379247Peptidase S8Add
    BLAST

    Sequence similaritiesi

    Belongs to the peptidase S8 family.Curated
    Contains 1 peptidase S8 domain.Curated

    Keywords - Domaini

    Signal

    Family and domain databases

    Gene3Di3.30.70.80. 1 hit.
    3.40.50.200. 1 hit.
    InterProiIPR000209. Peptidase_S8/S53_dom.
    IPR023827. Peptidase_S8_Asp-AS.
    IPR022398. Peptidase_S8_His-AS.
    IPR023828. Peptidase_S8_Ser-AS.
    IPR015500. Peptidase_S8_subtilisin-rel.
    IPR009020. Prot_inh_propept.
    IPR010259. S8pro/Inhibitor_I9.
    [Graphical view]
    PANTHERiPTHR10795. PTHR10795. 1 hit.
    PfamiPF05922. Inhibitor_I9. 1 hit.
    PF00082. Peptidase_S8. 1 hit.
    [Graphical view]
    PRINTSiPR00723. SUBTILISIN.
    SUPFAMiSSF52743. SSF52743. 1 hit.
    SSF54897. SSF54897. 1 hit.
    PROSITEiPS00136. SUBTILASE_ASP. 1 hit.
    PS00137. SUBTILASE_HIS. 1 hit.
    PS00138. SUBTILASE_SER. 1 hit.
    [Graphical view]

    Sequencei

    Sequence statusi: Complete.

    Sequence processingi: The displayed sequence is further processed into a mature form.

    P35835-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MRSKKLWISL LFALTLIFTM AFSNMSAQAA GKSSTEKKYI VGFKQTMSAM
    60 70 80 90 100
    SSAKKKDVIS EKGGKVQKQF KYVNAAAATL DEKAVKELKK DPSVAYVEED
    110 120 130 140 150
    HIAHEYAQSV PYGISQIKAP ALHSQGYTGS NVKVAVIDSG IDSSHPDLNV
    160 170 180 190 200
    RGGASFVPSE TNPYQDGSSH GTHVAGTIAA LNNSIGVLGV APSASLYAVK
    210 220 230 240 250
    VLDSTGSGQY SWIINGIEWA ISNNMDVINM SLGGPTGSTA LKTVVDKAVS
    260 270 280 290 300
    SGIVVAAAAG NEGSSGSTST VGYPAKYPST IAVGAVNSSN QRASFSSVGS
    310 320 330 340 350
    ELDVMAPGVS IQSTLPGGTY GAYNGTSMAT PHVAGAAALI LSKHPTWTNA
    360 370 380
    QVRDRLESTA TYLGNSFYYG KGLINVQAAA Q
    Length:381
    Mass (Da):39,507
    Last modified:June 1, 1994 - v1
    Checksum:iDAED4B16ED1BA092
    GO

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    D25319 Genomic DNA. Translation: BAA04989.1.
    S51909 Genomic DNA. Translation: AAC60424.1.
    PIRiJH0778.
    RefSeqiWP_014479360.1. NZ_CP014471.1.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    D25319 Genomic DNA. Translation: BAA04989.1.
    S51909 Genomic DNA. Translation: AAC60424.1.
    PIRiJH0778.
    RefSeqiWP_014479360.1. NZ_CP014471.1.

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    3VYVX-ray1.36A/B107-381[»]
    4DWWX-ray1.74A107-381[»]
    ProteinModelPortaliP35835.
    SMRiP35835. Positions 36-381.
    ModBaseiSearch...
    MobiDBiSearch...

    Protein family/group databases

    MEROPSiS08.044.

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Enzyme and pathway databases

    SABIO-RKP35835.

    Family and domain databases

    Gene3Di3.30.70.80. 1 hit.
    3.40.50.200. 1 hit.
    InterProiIPR000209. Peptidase_S8/S53_dom.
    IPR023827. Peptidase_S8_Asp-AS.
    IPR022398. Peptidase_S8_His-AS.
    IPR023828. Peptidase_S8_Ser-AS.
    IPR015500. Peptidase_S8_subtilisin-rel.
    IPR009020. Prot_inh_propept.
    IPR010259. S8pro/Inhibitor_I9.
    [Graphical view]
    PANTHERiPTHR10795. PTHR10795. 1 hit.
    PfamiPF05922. Inhibitor_I9. 1 hit.
    PF00082. Peptidase_S8. 1 hit.
    [Graphical view]
    PRINTSiPR00723. SUBTILISIN.
    SUPFAMiSSF52743. SSF52743. 1 hit.
    SSF54897. SSF54897. 1 hit.
    PROSITEiPS00136. SUBTILASE_ASP. 1 hit.
    PS00137. SUBTILASE_HIS. 1 hit.
    PS00138. SUBTILASE_SER. 1 hit.
    [Graphical view]
    ProtoNetiSearch...

    Publicationsi

    1. "Nucleotide sequence of the subtilisin NAT gene, aprN, of Bacillus subtilis (natto)."
      Nakamura T., Yamagata Y., Ichishima E.
      Biosci. Biotechnol. Biochem. 56:1869-1871(1992) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
      Strain: NC2-1.
    2. "Purification and characterization of a strong fibrinolytic enzyme (nattokinase) in the vegetable cheese natto, a popular soybean fermented food in Japan."
      Fujita M., Nomura K., Hong K., Ito Y., Asada A., Nishimuro S.
      Biochem. Biophys. Res. Commun. 197:1340-1347(1993) [PubMed] [Europe PMC] [Abstract]
      Cited for: PROTEIN SEQUENCE OF 107-381, CHARACTERIZATION.

    Entry informationi

    Entry nameiSUBN_BACNA
    AccessioniPrimary (citable) accession number: P35835
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: June 1, 1994
    Last sequence update: June 1, 1994
    Last modified: May 11, 2016
    This is version 95 of the entry and version 1 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    Miscellaneous

    Secretion of subtilisin is associated with onset of sporulation, and many mutations which block sporulation at early stages affect expression levels of subtilisin. However, subtilisin is not necessary for normal sporulation.
    In Japan this enzyme, isolated from a process of fermentation involving boiled soybeans and Bacillus natto, is used as a food supplement because it is reported to reduce and prevent blood clotting.

    Keywords - Technical termi

    3D-structure, Direct protein sequencing

    Documents

    1. PDB cross-references
      Index of Protein Data Bank (PDB) cross-references
    2. Peptidase families
      Classification of peptidase families and list of entries
    3. SIMILARITY comments
      Index of protein domains and families

    Similar proteinsi

    Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
    100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into one UniRef entry.
    90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
    50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.