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Protein

Subtilisin-like serine protease

Gene

TK1689

Organism
Thermococcus kodakarensis (strain ATCC BAA-918 / JCM 12380 / KOD1) (Pyrococcus kodakaraensis (strain KOD1))
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Serine protease with a broad substrate specificity.1 Publication

Catalytic activityi

Hydrolysis of proteins with broad specificity for peptide bonds, and a preference for a large uncharged residue in P1. Hydrolyzes peptide amides.1 Publication

Enzyme regulationi

Resistant to treatment with 5% SDS, 8 M urea, 10% Triton X-100 or 10% Tween-20. Fully active although less stable in the presence of 10 mM EDTA. Activity not affected by the absence or presence of 10 mM CaCl2. Unstable in the presence of 2 M or over GdnHCl and loses 35% and 99% of its activity upon incubation with 2 and 4 M GdnHCl, respectively, for 1 hour at 55 degrees Celsius. Nearly fully loses activity upon incubation at pH 2.0.1 Publication

Kineticsi

Kcat is 1.6 s(-1) for N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide at 20 degrees Celsius and pH 7.5(PubMed:20100702). Kcat is 25 s(-1) for N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide at 80 degrees Celsius and pH 7.5(PubMed:20100702).1 Publication

  1. KM=0.11 mM for N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide at 20 degrees Celsius and pH 7.51 Publication
  2. KM=0.41 mM for N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide at 80 degrees Celsius and pH 7.51 Publication
  1. Vmax=510 µmol/min/mg enzyme1 Publication

pH dependencei

High activity at a wide pH range between 7.0-11.5 at 20 degrees Celsius and pH 7.5. It is not fully stable at pH 6 or under and at pH 12 or over. It loses over 85% of its activity at pH 3 or under and at pH 13.1 Publication

Temperature dependencei

Optimum temperature is about 100 degrees Celsius. Highly thermostable. Stable at 80 degrees Celsius for at least 3 hours. Half-life at 100 degrees Celsius is 100 minutes and at 90 degrees Celsius more than 3 hours.1 Publication

Sites

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Active sitei170 – 1701Charge relay system1 Publication
Active sitei203 – 2031Charge relay system1 Publication
Sitei303 – 3031Important for catalytic activity1 Publication
Active sitei382 – 3821Nucleophile1 Publication
Metal bindingi420 – 4201Calcium 1; via carbonyl oxygen1 Publication
Metal bindingi423 – 4231Calcium 1; via carbonyl oxygen1 Publication
Metal bindingi483 – 4831Calcium 21 Publication
Metal bindingi484 – 4841Calcium 2; via carbonyl oxygen1 Publication
Metal bindingi485 – 4851Calcium 21 Publication
Metal bindingi497 – 4971Calcium 11 Publication
Metal bindingi498 – 4981Calcium 1; via carbonyl oxygen1 Publication
Metal bindingi501 – 5011Calcium 2; via carbonyl oxygen1 Publication
Metal bindingi507 – 5071Calcium 21 Publication

GO - Molecular functioni

  • calcium ion binding Source: UniProtKB
  • peptidase activity Source: UniProtKB
  • serine-type endopeptidase activity Source: UniProtKB

GO - Biological processi

  • proteolysis Source: UniProtKB
Complete GO annotation...

Keywords - Molecular functioni

Hydrolase, Protease, Serine protease

Keywords - Ligandi

Calcium, Metal-binding

Enzyme and pathway databases

BioCyciTKOD69014:GH72-1723-MONOMER.

Names & Taxonomyi

Protein namesi
Recommended name:
Subtilisin-like serine protease2 PublicationsImported (EC:3.4.21.621 Publication)
Alternative name(s):
Tk-SP2 Publications
Gene namesi
Ordered Locus Names:TK1689Imported
OrganismiThermococcus kodakarensis (strain ATCC BAA-918 / JCM 12380 / KOD1) (Pyrococcus kodakaraensis (strain KOD1))Imported
Taxonomic identifieri69014 [NCBI]
Taxonomic lineageiArchaeaEuryarchaeotaThermococciThermococcalesThermococcaceaeThermococcus
Proteomesi
  • UP000000536 Componenti: Chromosome

Pathology & Biotechi

Biotechnological usei

Has potential for application in biotechnology fields due to its high resistance to heat, denaturants, detergents and chelating agents.1 Publication

Mutagenesis

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Mutagenesisi382 – 3821S → A: Loss of activity. Greatly destabilized; when associated with the deletion of the calcium-binding form of the beta-jelly roll domain of the mature domain. 1 Publication
Mutagenesisi382 – 3821S → C: Greatly reduced enzymatic activity. Greatly reduced enzymatic activity; when associated with the deletion of the beta-jelly roll domain of the mature domain. 1 Publication

PTM / Processingi

Molecule processing

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Signal peptidei1 – 23231 PublicationAdd
BLAST
Propeptidei24 – 136113Removed in mature form2 PublicationsPRO_0000431236Add
BLAST
Chaini137 – 562426Subtilisin-like serine protease2 PublicationsPRO_0000431237Add
BLAST
Propeptidei563 – 663101Removed in mature form2 PublicationsPRO_0000431238Add
BLAST

Interactioni

Subunit structurei

Monomer.2 Publications

Protein-protein interaction databases

STRINGi69014.TK1689.

Structurei

Secondary structure

1
663
Legend: HelixTurnBeta strand
Show more details
Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Helixi44 – 518Combined sources
Beta strandi58 – 6710Combined sources
Helixi68 – 8114Combined sources
Beta strandi84 – 885Combined sources
Beta strandi90 – 10011Combined sources
Helixi101 – 1077Combined sources
Beta strandi124 – 1296Combined sources
Beta strandi132 – 1343Combined sources
Beta strandi165 – 1728Combined sources
Helixi178 – 1803Combined sources
Turni181 – 1833Combined sources
Beta strandi184 – 1896Combined sources
Turni190 – 1923Combined sources
Beta strandi199 – 2024Combined sources
Helixi203 – 21210Combined sources
Helixi216 – 2183Combined sources
Turni219 – 2224Combined sources
Beta strandi230 – 2356Combined sources
Beta strandi241 – 2444Combined sources
Helixi245 – 25713Combined sources
Helixi259 – 2624Combined sources
Beta strandi264 – 2696Combined sources
Helixi282 – 29211Combined sources
Beta strandi296 – 3005Combined sources
Beta strandi306 – 3094Combined sources
Turni313 – 3164Combined sources
Beta strandi318 – 3269Combined sources
Beta strandi334 – 3363Combined sources
Beta strandi349 – 3535Combined sources
Beta strandi355 – 3606Combined sources
Beta strandi368 – 3703Combined sources
Beta strandi372 – 3787Combined sources
Helixi381 – 39818Combined sources
Helixi404 – 41411Combined sources
Helixi420 – 4223Combined sources
Turni426 – 4283Combined sources
Helixi435 – 4395Combined sources
Helixi441 – 4433Combined sources
Beta strandi444 – 45310Combined sources
Beta strandi458 – 46710Combined sources
Beta strandi469 – 47810Combined sources
Beta strandi483 – 4897Combined sources
Beta strandi495 – 4995Combined sources
Beta strandi502 – 5054Combined sources
Beta strandi507 – 5137Combined sources
Beta strandi516 – 52813Combined sources
Beta strandi530 – 54314Combined sources

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
EntryMethodResolution (Å)ChainPositionsPDBsum
3AFGX-ray2.00A/B24-562[»]
ProteinModelPortaliQ5JIZ5.
ModBaseiSearch...
MobiDBiSearch...

Miscellaneous databases

EvolutionaryTraceiQ5JIZ5.

Family & Domainsi

Domains and Repeats

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Domaini165 – 439275Peptidase S8Sequence analysisAdd
BLAST

Domaini

The C-terminal calcium-binding beta-jelly roll domain (445-562) of the mature domain is not required for folding or activity, but it is required for the hyperstabilization of the protein and possibly for its adaptation to high-temperature environment.1 Publication

Sequence similaritiesi

Belongs to the peptidase S8 family.UniRule annotation
Contains 1 peptidase S8 domain.Sequence analysis

Keywords - Domaini

Signal

Phylogenomic databases

eggNOGiarCOG00702. Archaea.
COG1404. LUCA.
HOGENOMiHOG000199176.
KOiK17734.
OMAiVTATLYW.

Family and domain databases

Gene3Di3.40.50.200. 1 hit.
InterProiIPR008979. Galactose-bd-like.
IPR007280. Peptidase_C_arc/bac.
IPR000209. Peptidase_S8/S53_dom.
IPR023827. Peptidase_S8_Asp-AS.
IPR022398. Peptidase_S8_His-AS.
IPR023828. Peptidase_S8_Ser-AS.
IPR015500. Peptidase_S8_subtilisin-rel.
IPR017319. Subtilisin_TK1689.
[Graphical view]
PANTHERiPTHR10795. PTHR10795. 4 hits.
PfamiPF00082. Peptidase_S8. 1 hit.
PF04151. PPC. 2 hits.
[Graphical view]
PIRSFiPIRSF037907. Subtilisin_rel_TK1689. 1 hit.
PRINTSiPR00723. SUBTILISIN.
SUPFAMiSSF49785. SSF49785. 1 hit.
SSF52743. SSF52743. 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.

Q5JIZ5-1 [UniParc]FASTAAdd to basket

« Hide

        10         20         30         40         50
MKKFGAVVLA LFLVGLMAGS VLAAPQKPAV RNVSQQKNYG LLTPGLFKKV
60 70 80 90 100
QRMSWDQEVS TIIMFDNQAD KEKAVEILDF LGAKIKYNYH IIPALAVKIK
110 120 130 140 150
VKDLLIIAGL MDTGYFGNAQ LSGVQFIQED YVVKVAVETE GLDESAAQVM
160 170 180 190 200
ATNMWNLGYD GSGITIGIID TGIDASHPDL QGKVIGWVDF VNGKTTPYDD
210 220 230 240 250
NGHGTHVASI AAGTGAASNG KYKGMAPGAK LVGIKVLNGQ GSGSISDIIN
260 270 280 290 300
GVDWAVQNKD KYGIKVINLS LGSSQSSDGT DSLSQAVNNA WDAGLVVVVA
310 320 330 340 350
AGNSGPNKYT VGSPAAASKV ITVGAVDKYD VITDFSSRGP TADNRLKPEV
360 370 380 390 400
VAPGNWIIAA RASGTSMGQP INDYYTAAPG TSMATPHVAG IAALLLQAHP
410 420 430 440 450
SWTPDKVKTA LIETADIVKP DEIADIAYGA GRVNAYKAAY YDNYAKLTFT
460 470 480 490 500
GYVSNKGSQS HQFTISGAGF VTATLYWDNS GSDLDLYLYD PNGNQVDYSY
510 520 530 540 550
TAYYGFEKVG YYNPTAGTWT IKVVSYSGSA NYQVDVVSDG SLGQPSGGGS
560 570 580 590 600
EPSPSPSPEP TVDEKTFTGT VHDYYDKSDT FTMTVNSGAT KITGDLYFDT
610 620 630 640 650
SYHDLDLYLY DPNQNLVDRS ESSNSYEHVE YNNPAPGTWY FLVYAYDTYG
660
YADYQLDAKV YYG
Length:663
Mass (Da):70,955
Last modified:February 15, 2005 - v1
Checksum:i2CE68ACD3888E90E
GO

Mass spectrometryi

Molecular mass is 68600 Da from positions 24 - 663. Determined by MALDI. 1 Publication
Molecular mass is 44187±202 Da from positions 137 - 562. Determined by MALDI. 1 Publication

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
AP006878 Genomic DNA. Translation: BAD85878.1.
RefSeqiWP_011250640.1. NC_006624.1.

Genome annotation databases

EnsemblBacteriaiBAD85878; BAD85878; TK1689.
GeneIDi3234995.
KEGGitko:TK1689.

Cross-referencesi

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
AP006878 Genomic DNA. Translation: BAD85878.1.
RefSeqiWP_011250640.1. NC_006624.1.

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
EntryMethodResolution (Å)ChainPositionsPDBsum
3AFGX-ray2.00A/B24-562[»]
ProteinModelPortaliQ5JIZ5.
ModBaseiSearch...
MobiDBiSearch...

Protein-protein interaction databases

STRINGi69014.TK1689.

Protocols and materials databases

Structural Biology KnowledgebaseSearch...

Genome annotation databases

EnsemblBacteriaiBAD85878; BAD85878; TK1689.
GeneIDi3234995.
KEGGitko:TK1689.

Phylogenomic databases

eggNOGiarCOG00702. Archaea.
COG1404. LUCA.
HOGENOMiHOG000199176.
KOiK17734.
OMAiVTATLYW.

Enzyme and pathway databases

BioCyciTKOD69014:GH72-1723-MONOMER.

Miscellaneous databases

EvolutionaryTraceiQ5JIZ5.

Family and domain databases

Gene3Di3.40.50.200. 1 hit.
InterProiIPR008979. Galactose-bd-like.
IPR007280. Peptidase_C_arc/bac.
IPR000209. Peptidase_S8/S53_dom.
IPR023827. Peptidase_S8_Asp-AS.
IPR022398. Peptidase_S8_His-AS.
IPR023828. Peptidase_S8_Ser-AS.
IPR015500. Peptidase_S8_subtilisin-rel.
IPR017319. Subtilisin_TK1689.
[Graphical view]
PANTHERiPTHR10795. PTHR10795. 4 hits.
PfamiPF00082. Peptidase_S8. 1 hit.
PF04151. PPC. 2 hits.
[Graphical view]
PIRSFiPIRSF037907. Subtilisin_rel_TK1689. 1 hit.
PRINTSiPR00723. SUBTILISIN.
SUPFAMiSSF49785. SSF49785. 1 hit.
SSF52743. SSF52743. 1 hit.
PROSITEiPS00136. SUBTILASE_ASP. 1 hit.
PS00137. SUBTILASE_HIS. 1 hit.
PS00138. SUBTILASE_SER. 1 hit.
[Graphical view]
ProtoNetiSearch...

Publicationsi

« Hide 'large scale' publications
  1. "Complete genome sequence of the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1 and comparison with Pyrococcus genomes."
    Fukui T., Atomi H., Kanai T., Matsumi R., Fujiwara S., Imanaka T.
    Genome Res. 15:352-363(2005) [PubMed] [Europe PMC] [Abstract]
    Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
    Strain: ATCC BAA-918 / JCM 12380 / KOD1Imported.
  2. "Subtilisin-like serine protease from hyperthermophilic archaeon Thermococcus kodakaraensis with N- and C-terminal propeptides."
    Foophow T., Tanaka S., Koga Y., Takano K., Kanaya S.
    Protein Eng. Des. Sel. 23:347-355(2010) [PubMed] [Europe PMC] [Abstract]
    Cited for: PROTEIN SEQUENCE OF 24-26 AND 137-140, FUNCTION, CATALYTIC ACTIVITY, ENZYME REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, MASS SPECTROMETRY, BIOTECHNOLOGY, MUTAGENESIS OF SER-382, MAGNETIC CIRCULAR DICHROISM.
  3. "Crystal structure of a subtilisin homologue, Tk-SP, from Thermococcus kodakaraensis: requirement of a C-terminal beta-jelly roll domain for hyperstability."
    Foophow T., Tanaka S., Angkawidjaja C., Koga Y., Takano K., Kanaya S.
    J. Mol. Biol. 400:865-877(2010) [PubMed] [Europe PMC] [Abstract]
    Cited for: X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 24-562 OF MUTANT ALA-382 IN COMPLEX WITH CALCIUM, CATALYTIC ACTIVITY, SUBUNIT, DOMAIN, PRO-PEPTIDES, ACTIVE SITE, MUTAGENESIS OF SER-382, CIRCULAR DICHROISM.

Entry informationi

Entry nameiTKSP_THEKO
AccessioniPrimary (citable) accession number: Q5JIZ5
Entry historyi
Integrated into UniProtKB/Swiss-Prot: November 26, 2014
Last sequence update: February 15, 2005
Last modified: November 11, 2015
This is version 81 of the entry and version 1 of the sequence. [Complete history]
Entry statusiReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Miscellaneousi

Keywords - Technical termi

3D-structure, Complete proteome, Direct protein sequencing, Reference proteome

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.