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Protein

Subtilisin NAT

Gene

aprN

Organism
Bacillus subtilis subsp. natto
Status
Reviewed-Annotation score: -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.

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.

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)DescriptionActionsGraphical viewLength
    Metal bindingi108Calcium 1By similarity1
    Active sitei138Charge relay systemBy similarity1
    Metal bindingi147Calcium 1By similarity1
    Active sitei170Charge relay systemBy similarity1
    Metal bindingi181Calcium 1; via carbonyl oxygenBy similarity1
    Metal bindingi183Calcium 1By similarity1
    Metal bindingi185Calcium 1; via carbonyl oxygenBy similarity1
    Metal bindingi187Calcium 1; via carbonyl oxygenBy similarity1
    Metal bindingi275Calcium 2; via carbonyl oxygenBy similarity1
    Metal bindingi277Calcium 2; via carbonyl oxygenBy similarity1
    Metal bindingi280Calcium 2; via carbonyl oxygenBy similarity1
    Active sitei327Charge relay systemBy similarity1

    GO - Molecular functioni

    GO - Biological processi

    Keywordsi

    Molecular functionHydrolase, Protease, Serine protease
    Biological processBlood coagulation, Fibrinolysis, Hemostasis, Sporulation
    LigandCalcium, Metal-binding

    Enzyme and pathway databases

    SABIO-RKiP35835

    Protein family/group databases

    MEROPSiI09.001

    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

    Keywords - Cellular componenti

    Secreted

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)DescriptionActionsGraphical viewLength
    Signal peptidei1 – 29By similarityAdd BLAST29
    PropeptideiPRO_000002717530 – 1061 PublicationAdd BLAST77
    ChainiPRO_0000027176107 – 381Subtilisin NATAdd BLAST275

    Keywords - PTMi

    Zymogen

    Interactioni

    Subunit structurei

    Monomer.

    Structurei

    Secondary structure

    1381
    Legend: HelixTurnBeta strandPDB Structure known for this area
    Show more details
    Feature keyPosition(s)DescriptionActionsGraphical viewLength
    Helixi112 – 116Combined sources5
    Helixi119 – 125Combined sources7
    Beta strandi133 – 139Combined sources7
    Beta strandi150 – 155Combined sources6
    Beta strandi167 – 169Combined sources3
    Helixi170 – 179Combined sources10
    Beta strandi182 – 186Combined sources5
    Beta strandi195 – 200Combined sources6
    Helixi210 – 222Combined sources13
    Beta strandi226 – 230Combined sources5
    Helixi239 – 250Combined sources12
    Beta strandi254 – 258Combined sources5
    Turni274 – 276Combined sources3
    Beta strandi280 – 286Combined sources7
    Beta strandi304 – 307Combined sources4
    Beta strandi309 – 315Combined sources7
    Turni316 – 318Combined sources3
    Beta strandi319 – 323Combined sources5
    Helixi326 – 343Combined sources18
    Helixi349 – 358Combined sources10
    Helixi366 – 369Combined sources4
    Helixi376 – 379Combined sources4

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    PDB entryMethodResolution (Å)ChainPositionsPDBsum
    3VYVX-ray1.36A/B107-381[»]
    4DWWX-ray1.74A107-381[»]
    5GL8X-ray1.80A/B107-381[»]
    ProteinModelPortaliP35835
    SMRiP35835
    ModBaseiSearch...
    MobiDBiSearch...

    Family & Domainsi

    Domains and Repeats

    Feature keyPosition(s)DescriptionActionsGraphical viewLength
    Domaini133 – 379Peptidase S8Add BLAST247

    Sequence similaritiesi

    Belongs to the peptidase S8 family.Curated

    Keywords - Domaini

    Signal

    Family and domain databases

    CDDicd07477 Peptidases_S8_Subtilisin_subse, 1 hit
    Gene3Di3.30.70.80, 1 hit
    3.40.50.200, 1 hit
    InterProiView protein in InterPro
    IPR000209 Peptidase_S8/S53_dom
    IPR036852 Peptidase_S8/S53_dom_sf
    IPR023827 Peptidase_S8_Asp-AS
    IPR022398 Peptidase_S8_His-AS
    IPR023828 Peptidase_S8_Ser-AS
    IPR015500 Peptidase_S8_subtilisin-rel
    IPR010259 S8pro/Inhibitor_I9
    IPR037045 S8pro/Inhibitor_I9_sf
    IPR034202 Subtilisin_Carlsberg-like
    PfamiView protein in Pfam
    PF05922 Inhibitor_I9, 1 hit
    PF00082 Peptidase_S8, 1 hit
    PRINTSiPR00723 SUBTILISIN
    SUPFAMiSSF52743 SSF52743, 1 hit
    PROSITEiView protein in PROSITE
    PS00136 SUBTILASE_ASP, 1 hit
    PS00137 SUBTILASE_HIS, 1 hit
    PS00138 SUBTILASE_SER, 1 hit

    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

    Similar proteinsi

    Entry informationi

    Entry nameiSUBN_BACNA
    AccessioniPrimary (citable) accession number: P35835
    Entry historyiIntegrated into UniProtKB/Swiss-Prot: June 1, 1994
    Last sequence update: June 1, 1994
    Last modified: April 25, 2018
    This is version 107 of the entry and version 1 of the sequence. See complete history.
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    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

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