TKSP_THEKO
ID TKSP_THEKO Reviewed; 663 AA.
AC Q5JIZ5;
DT 26-NOV-2014, integrated into UniProtKB/Swiss-Prot.
DT 15-FEB-2005, sequence version 1.
DT 03-AUG-2022, entry version 107.
DE RecName: Full=Subtilisin-like serine protease {ECO:0000303|PubMed:20100702, ECO:0000303|PubMed:20595040, ECO:0000312|EMBL:BAD85878.1};
DE EC=3.4.21.62 {ECO:0000269|PubMed:20100702};
DE AltName: Full=Tk-SP {ECO:0000303|PubMed:20100702, ECO:0000303|PubMed:20595040};
DE Flags: Precursor;
GN OrderedLocusNames=TK1689 {ECO:0000312|EMBL:BAD85878.1};
OS Thermococcus kodakarensis (strain ATCC BAA-918 / JCM 12380 / KOD1)
OS (Pyrococcus kodakaraensis (strain KOD1)).
OC Archaea; Euryarchaeota; Thermococci; Thermococcales; Thermococcaceae;
OC Thermococcus.
OX NCBI_TaxID=69014 {ECO:0000312|Proteomes:UP000000536};
RN [1] {ECO:0000312|EMBL:BAD85878.1, ECO:0000312|Proteomes:UP000000536}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC BAA-918 / JCM 12380 / KOD1 {ECO:0000312|Proteomes:UP000000536};
RX PubMed=15710748; DOI=10.1101/gr.3003105;
RA Fukui T., Atomi H., Kanai T., Matsumi R., Fujiwara S., Imanaka T.;
RT "Complete genome sequence of the hyperthermophilic archaeon Thermococcus
RT kodakaraensis KOD1 and comparison with Pyrococcus genomes.";
RL Genome Res. 15:352-363(2005).
RN [2]
RP PROTEIN SEQUENCE OF 24-26 AND 137-140, FUNCTION, CATALYTIC ACTIVITY,
RP ACTIVITY REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, MASS
RP SPECTROMETRY, BIOTECHNOLOGY, MUTAGENESIS OF SER-382, AND MAGNETIC CIRCULAR
RP DICHROISM.
RX PubMed=20100702; DOI=10.1093/protein/gzp092;
RA Foophow T., Tanaka S., Koga Y., Takano K., Kanaya S.;
RT "Subtilisin-like serine protease from hyperthermophilic archaeon
RT Thermococcus kodakaraensis with N- and C-terminal propeptides.";
RL Protein Eng. Des. Sel. 23:347-355(2010).
RN [3] {ECO:0000312|PDB:3AFG}
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 24-562 OF MUTANT ALA-382 IN
RP COMPLEX WITH CALCIUM, CATALYTIC ACTIVITY, SUBUNIT, DOMAIN, PRO-PEPTIDES,
RP ACTIVE SITE, MUTAGENESIS OF SER-382, AND CIRCULAR DICHROISM.
RX PubMed=20595040; DOI=10.1016/j.jmb.2010.05.064;
RA Foophow T., Tanaka S., Angkawidjaja C., Koga Y., Takano K., Kanaya S.;
RT "Crystal structure of a subtilisin homologue, Tk-SP, from Thermococcus
RT kodakaraensis: requirement of a C-terminal beta-jelly roll domain for
RT hyperstability.";
RL J. Mol. Biol. 400:865-877(2010).
CC -!- FUNCTION: Serine protease with a broad substrate specificity.
CC {ECO:0000269|PubMed:20100702}.
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:20100702};
CC -!- ACTIVITY REGULATION: Resistant to treatment with 5% SDS, 8 M urea, 10%
CC Triton X-100 or 10% Tween-20. Fully active although less stable in the
CC presence of 10 mM EDTA. Activity not affected by the absence or
CC presence of 10 mM CaCl(2). Unstable in the presence of 2 M or over
CC GdnHCl and loses 35% and 99% of its activity upon incubation with 2 and
CC 4 M GdnHCl, respectively, for 1 hour at 55 degrees Celsius. Nearly
CC fully loses activity upon incubation at pH 2.0.
CC {ECO:0000269|PubMed:20100702}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.11 mM for N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide at 20
CC degrees Celsius and pH 7.5 {ECO:0000269|PubMed:20100702};
CC KM=0.41 mM for N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide at 80
CC degrees Celsius and pH 7.5 {ECO:0000269|PubMed:20100702};
CC Vmax=510 umol/min/mg enzyme {ECO:0000269|PubMed:20100702};
CC Note=kcat is 1.6 sec(-1) for N-succinyl-Ala-Ala-Pro-Phe-p-
CC nitroanilide at 20 degrees Celsius and pH 7.5(PubMed:20100702). kcat
CC is 25 sec(-1) for N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide at 80
CC degrees Celsius and pH 7.5(PubMed:20100702).
CC {ECO:0000269|PubMed:20100702};
CC pH dependence:
CC High activity at a wide pH range between 7.0-11.5 at 20 degrees
CC Celsius and pH 7.5. It is not fully stable at pH 6 or under and at pH
CC 12 or over. It loses over 85% of its activity at pH 3 or under and at
CC pH 13. {ECO:0000269|PubMed:20100702};
CC Temperature dependence:
CC Optimum temperature is about 100 degrees Celsius. Highly
CC thermostable. Stable at 80 degrees Celsius for at least 3 hours.
CC Half-life at 100 degrees Celsius is 100 minutes and at 90 degrees
CC Celsius more than 3 hours. {ECO:0000269|PubMed:20100702};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:20100702,
CC ECO:0000269|PubMed:20595040}.
CC -!- DOMAIN: The C-terminal calcium-binding beta-jelly roll domain (445-562)
CC of the mature domain is not required for folding or activity, but it is
CC required for the hyperstabilization of the protein and possibly for its
CC adaptation to high-temperature environment.
CC {ECO:0000269|PubMed:20595040}.
CC -!- MASS SPECTROMETRY: Mass=44187; Mass_error=202; Method=MALDI;
CC Evidence={ECO:0000269|PubMed:20100702};
CC -!- BIOTECHNOLOGY: Has potential for application in biotechnology fields
CC due to its high resistance to heat, denaturants, detergents and
CC chelating agents. {ECO:0000303|PubMed:20100702}.
CC -!- SIMILARITY: Belongs to the peptidase S8 family.
CC {ECO:0000255|RuleBase:RU003355}.
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DR EMBL; AP006878; BAD85878.1; -; Genomic_DNA.
DR RefSeq; WP_011250640.1; NC_006624.1.
DR PDB; 3AFG; X-ray; 2.00 A; A/B=24-562.
DR PDBsum; 3AFG; -.
DR AlphaFoldDB; Q5JIZ5; -.
DR SMR; Q5JIZ5; -.
DR STRING; 69014.TK1689; -.
DR EnsemblBacteria; BAD85878; BAD85878; TK1689.
DR GeneID; 3234995; -.
DR KEGG; tko:TK1689; -.
DR PATRIC; fig|69014.16.peg.1647; -.
DR eggNOG; arCOG00702; Archaea.
DR HOGENOM; CLU_011263_15_5_2; -.
DR InParanoid; Q5JIZ5; -.
DR OMA; ATLYWDN; -.
DR OrthoDB; 53822at2157; -.
DR PhylomeDB; Q5JIZ5; -.
DR BRENDA; 3.4.21.62; 5246.
DR EvolutionaryTrace; Q5JIZ5; -.
DR Proteomes; UP000000536; Chromosome.
DR GO; GO:0005509; F:calcium ion binding; IDA:UniProtKB.
DR GO; GO:0008233; F:peptidase activity; IDA:UniProtKB.
DR GO; GO:0004252; F:serine-type endopeptidase activity; IDA:UniProtKB.
DR GO; GO:0006508; P:proteolysis; IDA:UniProtKB.
DR Gene3D; 3.30.70.80; -; 1.
DR Gene3D; 3.40.50.200; -; 1.
DR InterPro; IPR008979; Galactose-bd-like_sf.
DR InterPro; IPR007280; Peptidase_C_arc/bac.
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; IPR037045; S8pro/Inhibitor_I9_sf.
DR InterPro; IPR017319; Subtilisin_TK1689.
DR InterPro; IPR041326; Tk-SP_N-pro.
DR Pfam; PF00082; Peptidase_S8; 1.
DR Pfam; PF04151; PPC; 1.
DR Pfam; PF18237; Tk-SP_N-pro; 1.
DR PIRSF; PIRSF037907; Subtilisin_rel_TK1689; 1.
DR PRINTS; PR00723; SUBTILISIN.
DR SUPFAM; SSF49785; SSF49785; 1.
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; Reference proteome; Serine protease; Signal.
FT SIGNAL 1..23
FT /evidence="ECO:0000269|PubMed:20595040"
FT PROPEP 24..136
FT /note="Removed in mature form"
FT /evidence="ECO:0000269|PubMed:20100702,
FT ECO:0000269|PubMed:20595040"
FT /id="PRO_0000431236"
FT CHAIN 137..562
FT /note="Subtilisin-like serine protease"
FT /evidence="ECO:0000269|PubMed:20100702,
FT ECO:0000269|PubMed:20595040"
FT /id="PRO_0000431237"
FT PROPEP 563..663
FT /note="Removed in mature form"
FT /evidence="ECO:0000269|PubMed:20100702,
FT ECO:0000269|PubMed:20595040"
FT /id="PRO_0000431238"
FT DOMAIN 139..439
FT /note="Peptidase S8"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240"
FT REGION 537..565
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 170
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240,
FT ECO:0000269|PubMed:20595040"
FT ACT_SITE 203
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240,
FT ECO:0000269|PubMed:20595040"
FT ACT_SITE 382
FT /note="Charge relay system"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01240,
FT ECO:0000269|PubMed:20595040"
FT BINDING 420
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:20595040"
FT BINDING 423
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:20595040"
FT BINDING 483
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:20595040"
FT BINDING 484
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:20595040"
FT BINDING 485
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:20595040"
FT BINDING 497
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:20595040"
FT BINDING 498
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:20595040"
FT BINDING 501
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:20595040"
FT BINDING 507
FT /ligand="Ca(2+)"
FT /ligand_id="ChEBI:CHEBI:29108"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:20595040"
FT SITE 303
FT /note="Important for catalytic activity"
FT /evidence="ECO:0000303|PubMed:20100702"
FT MUTAGEN 382
FT /note="S->A: Loss of activity. Greatly destabilized; when
FT associated with the deletion of the calcium-binding form of
FT the beta-jelly roll domain of the mature domain."
FT /evidence="ECO:0000269|PubMed:20595040"
FT MUTAGEN 382
FT /note="S->C: Greatly reduced enzymatic activity. Greatly
FT reduced enzymatic activity; when associated with the
FT deletion of the beta-jelly roll domain of the mature
FT domain."
FT /evidence="ECO:0000269|PubMed:20595040"
FT HELIX 44..51
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 58..67
FT /evidence="ECO:0007829|PDB:3AFG"
FT HELIX 68..81
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 84..88
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 90..100
FT /evidence="ECO:0007829|PDB:3AFG"
FT HELIX 101..107
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 124..129
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 132..134
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 165..172
FT /evidence="ECO:0007829|PDB:3AFG"
FT HELIX 178..180
FT /evidence="ECO:0007829|PDB:3AFG"
FT TURN 181..183
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 184..189
FT /evidence="ECO:0007829|PDB:3AFG"
FT TURN 190..192
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 199..202
FT /evidence="ECO:0007829|PDB:3AFG"
FT HELIX 203..212
FT /evidence="ECO:0007829|PDB:3AFG"
FT HELIX 216..218
FT /evidence="ECO:0007829|PDB:3AFG"
FT TURN 219..222
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 230..235
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 241..244
FT /evidence="ECO:0007829|PDB:3AFG"
FT HELIX 245..257
FT /evidence="ECO:0007829|PDB:3AFG"
FT HELIX 259..262
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 264..269
FT /evidence="ECO:0007829|PDB:3AFG"
FT HELIX 282..292
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 296..300
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 306..309
FT /evidence="ECO:0007829|PDB:3AFG"
FT TURN 313..316
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 318..326
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 334..336
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 349..353
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 355..360
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 368..370
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 372..378
FT /evidence="ECO:0007829|PDB:3AFG"
FT HELIX 381..398
FT /evidence="ECO:0007829|PDB:3AFG"
FT HELIX 404..414
FT /evidence="ECO:0007829|PDB:3AFG"
FT HELIX 420..422
FT /evidence="ECO:0007829|PDB:3AFG"
FT TURN 426..428
FT /evidence="ECO:0007829|PDB:3AFG"
FT HELIX 435..439
FT /evidence="ECO:0007829|PDB:3AFG"
FT HELIX 441..443
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 444..453
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 458..467
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 469..478
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 483..489
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 495..499
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 502..505
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 507..513
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 516..528
FT /evidence="ECO:0007829|PDB:3AFG"
FT STRAND 530..543
FT /evidence="ECO:0007829|PDB:3AFG"
SQ SEQUENCE 663 AA; 70955 MW; 2CE68ACD3888E90E CRC64;
MKKFGAVVLA LFLVGLMAGS VLAAPQKPAV RNVSQQKNYG LLTPGLFKKV QRMSWDQEVS
TIIMFDNQAD KEKAVEILDF LGAKIKYNYH IIPALAVKIK VKDLLIIAGL MDTGYFGNAQ
LSGVQFIQED YVVKVAVETE GLDESAAQVM ATNMWNLGYD GSGITIGIID TGIDASHPDL
QGKVIGWVDF VNGKTTPYDD NGHGTHVASI AAGTGAASNG KYKGMAPGAK LVGIKVLNGQ
GSGSISDIIN GVDWAVQNKD KYGIKVINLS LGSSQSSDGT DSLSQAVNNA WDAGLVVVVA
AGNSGPNKYT VGSPAAASKV ITVGAVDKYD VITDFSSRGP TADNRLKPEV VAPGNWIIAA
RASGTSMGQP INDYYTAAPG TSMATPHVAG IAALLLQAHP SWTPDKVKTA LIETADIVKP
DEIADIAYGA GRVNAYKAAY YDNYAKLTFT GYVSNKGSQS HQFTISGAGF VTATLYWDNS
GSDLDLYLYD PNGNQVDYSY TAYYGFEKVG YYNPTAGTWT IKVVSYSGSA NYQVDVVSDG
SLGQPSGGGS EPSPSPSPEP TVDEKTFTGT VHDYYDKSDT FTMTVNSGAT KITGDLYFDT
SYHDLDLYLY DPNQNLVDRS ESSNSYEHVE YNNPAPGTWY FLVYAYDTYG YADYQLDAKV
YYG
