SUBC_BACLI
ID SUBC_BACLI Reviewed; 379 AA.
AC P00780; Q9F943;
DT 21-JUL-1986, integrated into UniProtKB/Swiss-Prot.
DT 18-SEP-2019, sequence version 2.
DT 03-AUG-2022, entry version 164.
DE RecName: Full=Subtilisin Carlsberg {ECO:0000303|PubMed:4967581};
DE EC=3.4.21.62 {ECO:0000269|PubMed:11109488, ECO:0000269|Ref.4};
DE Flags: Precursor;
GN Name=subC {ECO:0000312|EMBL:CAB56500.1}; Synonyms=apr;
OS Bacillus licheniformis.
OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX NCBI_TaxID=1402;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=NCIMB 6816 / CCM 2182 / NCDO 727 / NCTC 6816;
RX PubMed=3001653; DOI=10.1093/nar/13.24.8913;
RA Jacobs M., Eliasson M., Uhlen M., Flock J.-I.;
RT "Cloning, sequencing and expression of subtilisin Carlsberg from Bacillus
RT licheniformis.";
RL Nucleic Acids Res. 13:8913-8926(1985).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-374, FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND SUBCELLULAR LOCATION.
RC STRAIN=NCIMB 6816 / CCM 2182 / NCDO 727 / NCTC 6816;
RX PubMed=11109488; DOI=10.1139/w00-085;
RA Evans K.L., Crowder J., Miller E.S.;
RT "Subtilisins of Bacillus spp. hydrolyze keratin and allow growth on
RT feathers.";
RL Can. J. Microbiol. 46:1004-1011(2000).
RN [3]
RP PROTEIN SEQUENCE OF 106-379.
RX PubMed=4967581; DOI=10.1016/s0021-9258(18)93461-7;
RA Smith E.L., Delange R.J., Evans W.H., Landon M., Markland F.S.;
RT "Subtilisin Carlsberg. V. The complete sequence; comparison with subtilisin
RT BPN'; evolutionary relationships.";
RL J. Biol. Chem. 243:2184-2191(1968).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX DOI=10.1007/BF01569566;
RA Peters K., Pauli D., Hache H., Boteva R.N., Genov N.C., Fittkau S.;
RT "Subtilisin DY-Kinetic characterization and comparison with related
RT proteinases.";
RL Curr. Microbiol. 18:171-177(1989).
RN [5] {ECO:0007744|PDB:1SEL}
RP X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) OF 106-379 OF MUTANT
RP SELENOCYSTEINE-325 IN COMPLEX WITH CALCIUM, AND COFACTOR.
RX PubMed=8512925; DOI=10.1021/bi00075a007;
RA Syed R., Wu Z.P., Hogle J.M., Hilvert D.;
RT "Crystal structure of selenosubtilisin at 2.0-A resolution.";
RL Biochemistry 32:6157-6164(1993).
RN [6] {ECO:0007744|PDB:1AV7, ECO:0007744|PDB:1AVT, ECO:0007744|PDB:1VSB, ECO:0007744|PDB:3VSB}
RP X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS) OF 106-379, AND ACTIVITY REGULATION.
RX PubMed=9425066; DOI=10.1021/bi971166o;
RA Stoll V.S., Eger B.T., Hynes R.C., Martichonok V., Jones J.B., Pai E.F.;
RT "Differences in binding modes of enantiomers of 1-acetamido boronic acid
RT based protease inhibitors: crystal structures of gamma-chymotrypsin and
RT subtilisin Carlsberg complexes.";
RL Biochemistry 37:451-462(1998).
CC -!- FUNCTION: Subtilisin is an extracellular alkaline serine protease, it
CC catalyzes the hydrolysis of proteins and peptide amides (Ref.4,
CC PubMed:11109488). Shows high specificity for aromatic and hydrophobic
CC amino acids in the P1 substrate position (PubMed:11109488). May play an
CC important role in the degradation of feather keratin (PubMed:11109488).
CC {ECO:0000269|PubMed:11109488, ECO:0000269|Ref.4}.
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 Evidence={ECO:0000269|PubMed:11109488, ECO:0000269|Ref.4};
CC -!- COFACTOR:
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC Evidence={ECO:0000269|PubMed:8512925};
CC Note=Binds 2 calcium ions per subunit. {ECO:0000269|PubMed:8512925};
CC -!- ACTIVITY REGULATION: Inhibited by p-chlorophenyl and 1-naphthyl boronic
CC acid derivatives. {ECO:0000269|PubMed:9425066}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.2 mM for Suc-Ala(2)-Ala-pNA {ECO:0000269|Ref.4};
CC KM=0.79 mM for Suc-Ala(2)-Phe-pNA {ECO:0000269|Ref.4};
CC KM=0.851 mM for Suc-Ala(2)-Phe-pNA {ECO:0000269|PubMed:11109488};
CC KM=0.22 mM for MeO-Suc-Ala(2)-Phe-pNA {ECO:0000269|Ref.4};
CC KM=5.6 mM for Suc-Gly(2)-Phe-pNA {ECO:0000269|PubMed:11109488};
CC KM=0.334 mM for Suc-Ala(2)-Pro-Phe-pNA {ECO:0000269|PubMed:11109488};
CC KM=0.062 mM for Suc-Ala(2)-Pro-Leu-pNA {ECO:0000269|PubMed:11109488};
CC KM=0.422 mM for Suc-Ala(2)-Pro-Arg-pNA {ECO:0000269|PubMed:11109488};
CC KM=0.245 mM for Suc-Ala(2)-Pro-Glu-pNA {ECO:0000269|PubMed:11109488};
CC KM=0.042 mM for Suc-Ala-Glu-Pro-Phe-pNA
CC {ECO:0000269|PubMed:11109488};
CC Note=kcat is 1.6 sec(-1) with Suc-Ala(2)-Ala-pNA as substrate. kcat
CC is 57 sec(-1) with Suc-Ala(2)-Phe-pNA as substrate. kcat is 97 sec(-
CC 1) with MeO-Suc-Ala(2)-Phe-pNA as substrate (Ref.4). kcat is 0.990
CC sec(-1) with Suc-Gly(2)-Phe-pNA as substrate. kcat is 20.3 sec(-1)
CC with Suc-Ala(2)-Phe-pNA as substrate. kcat is 646 sec(-1) with Suc-
CC Ala(2)-Pro-Phe-pNA as substrate. kcat is 137 sec(-1) with Suc-Ala(2)-
CC Pro-Leu-pNA as substrate. kcat is 5.31 sec(-1) with Suc-Ala(2)-Pro-
CC Arg-pNA as substrate. kcat is 1.92 sec(-1) with Suc-Ala(2)-Pro-Glu-
CC pNA as substrate. kcat is 375 sec(-1) with Suc-Ala-Glu-Pro-Phe-pNA as
CC substrate (PubMed:11109488). {ECO:0000269|PubMed:11109488,
CC ECO:0000269|Ref.4};
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000269|PubMed:11109488}.
CC -!- BIOTECHNOLOGY: Used as a detergent protease. Sold under the name
CC Alcalase by Novozymes.
CC -!- SIMILARITY: Belongs to the peptidase S8 family. {ECO:0000255|PROSITE-
CC ProRule:PRU01240}.
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DR EMBL; X03341; CAB56500.1; -; Genomic_DNA.
DR EMBL; AF205189; AAG31026.1; -; Genomic_DNA.
DR PIR; A24111; SUBSCL.
DR PDB; 1AF4; X-ray; 2.60 A; A=106-379.
DR PDB; 1AV7; X-ray; 2.60 A; A=106-379.
DR PDB; 1AVT; X-ray; 2.00 A; A=106-379.
DR PDB; 1BE6; X-ray; 2.15 A; A=106-379.
DR PDB; 1BE8; X-ray; 2.20 A; A=106-379.
DR PDB; 1BFK; X-ray; 2.30 A; A=106-379.
DR PDB; 1BFU; X-ray; 2.20 A; A=106-379.
DR PDB; 1C3L; X-ray; 2.16 A; A=106-378.
DR PDB; 1CSE; X-ray; 1.20 A; E=106-379.
DR PDB; 1OYV; X-ray; 2.50 A; A/B=106-379.
DR PDB; 1R0R; X-ray; 1.10 A; E=106-378.
DR PDB; 1SBC; X-ray; 2.50 A; A=106-379.
DR PDB; 1SCA; X-ray; 2.00 A; A=106-379.
DR PDB; 1SCB; X-ray; 2.30 A; A=106-379.
DR PDB; 1SCD; X-ray; 2.30 A; A=106-379.
DR PDB; 1SCN; X-ray; 1.90 A; E=106-379.
DR PDB; 1SEL; X-ray; 2.00 A; A/B=106-379.
DR PDB; 1VSB; X-ray; 2.10 A; A=106-379.
DR PDB; 1YU6; X-ray; 1.55 A; A/B=106-379.
DR PDB; 2SEC; X-ray; 1.80 A; E=106-379.
DR PDB; 2WUV; X-ray; 2.24 A; A=106-379.
DR PDB; 2WUW; X-ray; 2.23 A; E=106-379.
DR PDB; 3UNX; X-ray; 1.26 A; A=106-379.
DR PDB; 3VSB; X-ray; 2.60 A; A=106-379.
DR PDB; 4C3U; X-ray; 2.29 A; A=106-379.
DR PDB; 4C3V; X-ray; 2.26 A; A=106-379.
DR PDB; 6DWQ; X-ray; 1.27 A; A=106-379.
DR PDBsum; 1AF4; -.
DR PDBsum; 1AV7; -.
DR PDBsum; 1AVT; -.
DR PDBsum; 1BE6; -.
DR PDBsum; 1BE8; -.
DR PDBsum; 1BFK; -.
DR PDBsum; 1BFU; -.
DR PDBsum; 1C3L; -.
DR PDBsum; 1CSE; -.
DR PDBsum; 1OYV; -.
DR PDBsum; 1R0R; -.
DR PDBsum; 1SBC; -.
DR PDBsum; 1SCA; -.
DR PDBsum; 1SCB; -.
DR PDBsum; 1SCD; -.
DR PDBsum; 1SCN; -.
DR PDBsum; 1SEL; -.
DR PDBsum; 1VSB; -.
DR PDBsum; 1YU6; -.
DR PDBsum; 2SEC; -.
DR PDBsum; 2WUV; -.
DR PDBsum; 2WUW; -.
DR PDBsum; 3UNX; -.
DR PDBsum; 3VSB; -.
DR PDBsum; 4C3U; -.
DR PDBsum; 4C3V; -.
DR PDBsum; 6DWQ; -.
DR AlphaFoldDB; P00780; -.
DR PCDDB; P00780; -.
DR SMR; P00780; -.
DR IntAct; P00780; 2.
DR MINT; P00780; -.
DR BindingDB; P00780; -.
DR ChEMBL; CHEMBL4299; -.
DR DrugBank; DB03316; 1,4-Dioxane.
DR DrugBank; DB03607; [(1R)-1-acetamido-2-(4-chlorophenyl)ethyl]-[(2S)-2-amino-3-hydroxy-3-oxo-propoxy]-dihydroxy-boron.
DR DrugBank; DB02560; [(1S)-1-acetamido-2-(4-chlorophenyl)ethyl]-[(2S)-2-amino-3-hydroxy-3-oxo-propoxy]-dihydroxy-boron.
DR DrugBank; DB02677; D-naphthyl-1-acetamido boronic acid alanine.
DR DrugBank; DB01942; Formic acid.
DR DrugBank; DB03290; L-naphthyl-1-acetamido boronic acid alanine.
DR Allergome; 8254; Bac li Subtilisin.
DR MEROPS; I09.001; -.
DR MEROPS; S08.001; -.
DR MetOSite; P00780; -.
DR PRIDE; P00780; -.
DR BRENDA; 3.4.21.62; 669.
DR SABIO-RK; P00780; -.
DR EvolutionaryTrace; P00780; -.
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:0006508; P:proteolysis; 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; Calcium; Direct protein sequencing; Hydrolase; Metal-binding;
KW Protease; Secreted; Serine protease; Signal; Zymogen.
FT SIGNAL 1..29
FT /evidence="ECO:0000255"
FT PROPEP 30..105
FT /evidence="ECO:0000269|PubMed:4967581"
FT /id="PRO_0000027179"
FT CHAIN 106..379
FT /note="Subtilisin Carlsberg"
FT /evidence="ECO:0000269|PubMed:4967581"
FT /id="PRO_0000027180"
FT DOMAIN 44..102
FT /note="Inhibitor I9"
FT /evidence="ECO:0000255"
FT DOMAIN 110..378
FT /note="Peptidase S8"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT ACT_SITE 137
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT ACT_SITE 168
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT ACT_SITE 325
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT BINDING 107
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:8512925"
FT BINDING 146
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:8512925"
FT BINDING 179
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:8512925"
FT BINDING 181
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:8512925"
FT BINDING 183
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:8512925"
FT BINDING 185
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:8512925"
FT BINDING 273
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:8512925"
FT BINDING 275
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:8512925"
FT BINDING 278
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:8512925"
FT CONFLICT 207
FT /note="S -> T (in Ref. 1; CAB56500)"
FT /evidence="ECO:0000305"
FT CONFLICT 233
FT /note="A -> P (in Ref. 1; CAB56500)"
FT /evidence="ECO:0000305"
FT CONFLICT 249
FT /note="K -> R (in Ref. 1; CAB56500 and 3; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 262..265
FT /note="SSGN -> NSGS (in Ref. 3; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 316
FT /note="N -> S (in Ref. 1; CAB56500)"
FT /evidence="ECO:0000305"
FT HELIX 112..115
FT /evidence="ECO:0007829|PDB:1R0R"
FT HELIX 118..123
FT /evidence="ECO:0007829|PDB:1R0R"
FT STRAND 132..138
FT /evidence="ECO:0007829|PDB:1R0R"
FT STRAND 149..154
FT /evidence="ECO:0007829|PDB:1R0R"
FT STRAND 156..158
FT /evidence="ECO:0007829|PDB:1SBC"
FT STRAND 161..163
FT /evidence="ECO:0007829|PDB:1SBC"
FT STRAND 165..167
FT /evidence="ECO:0007829|PDB:1R0R"
FT HELIX 168..177
FT /evidence="ECO:0007829|PDB:1R0R"
FT STRAND 180..184
FT /evidence="ECO:0007829|PDB:1R0R"
FT STRAND 192..198
FT /evidence="ECO:0007829|PDB:1R0R"
FT STRAND 204..206
FT /evidence="ECO:0007829|PDB:1R0R"
FT HELIX 208..220
FT /evidence="ECO:0007829|PDB:1R0R"
FT STRAND 224..228
FT /evidence="ECO:0007829|PDB:1R0R"
FT STRAND 230..234
FT /evidence="ECO:0007829|PDB:1R0R"
FT HELIX 237..248
FT /evidence="ECO:0007829|PDB:1R0R"
FT STRAND 252..256
FT /evidence="ECO:0007829|PDB:1R0R"
FT TURN 272..274
FT /evidence="ECO:0007829|PDB:1R0R"
FT STRAND 278..284
FT /evidence="ECO:0007829|PDB:1R0R"
FT STRAND 300..305
FT /evidence="ECO:0007829|PDB:1R0R"
FT STRAND 307..313
FT /evidence="ECO:0007829|PDB:1R0R"
FT TURN 314..316
FT /evidence="ECO:0007829|PDB:1R0R"
FT STRAND 317..321
FT /evidence="ECO:0007829|PDB:1R0R"
FT HELIX 324..341
FT /evidence="ECO:0007829|PDB:1R0R"
FT HELIX 347..356
FT /evidence="ECO:0007829|PDB:1R0R"
FT HELIX 364..367
FT /evidence="ECO:0007829|PDB:1R0R"
FT HELIX 374..377
FT /evidence="ECO:0007829|PDB:1R0R"
SQ SEQUENCE 379 AA; 38867 MW; 38432C04FFA048C1 CRC64;
MMRKKSFWLG MLTAFMLVFT MAFSDSASAA QPAKNVEKDY IVGFKSGVKT ASVKKDIIKE
SGGKVDKQFR IINAAKAKLD KEALKEVKND PDVAYVEEDH VAHALAQTVP YGIPLIKADK
VQAQGFKGAN VKVAVLDTGI QASHPDLNVV GGASFVAGEA YNTDGNGHGT HVAGTVAALD
NTTGVLGVAP SVSLYAVKVL NSSGSGSYSG IVSGIEWATT NGMDVINMSL GGASGSTAMK
QAVDNAYAKG VVVVAAAGNS GSSGNTNTIG YPAKYDSVIA VGAVDSNSNR ASFSSVGAEL
EVMAPGAGVY STYPTNTYAT LNGTSMASPH VAGAAALILS KHPNLSASQV RNRLSSTATY
LGSSFYYGKG LINVEAAAQ
