EzCatDB: S00519
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DB codeS00519
RLCP classification1.13.30000.9 : Hydrolysis
CATH domainDomain 13.40.50.200 : Rossmann foldCatalytic domain
E.C.3.4.21.66

CATH domainRelated DB codes (homologues)
3.40.50.200 : Rossmann foldS00295,S00296,D00219

Enzyme Name
UniProtKBKEGG

P04072
Protein nameThermitasethermitase
thermophilic Streptomyces serine proteinase
Thermoactinomyces vulgaris serine proteinase
SynonymsEC 3.4.21.66
MEROPSS08.007 (Serine)
PfamPF00082 (Peptidase_S8)
[Graphical view]


UniProtKB:Accession NumberP04072
Entry nameTHET_THEVU
ActivityHydrolysis of proteins, including collagen.
Subunit
Subcellular locationSecreted.
CofactorBinds 3 calcium ions or 2 calcium ions and 1 sodium ion per subunit. The sodium ion is bound at calcium concentrations up to 5 mM. At 100 mM calcium 3 calcium ions are bound.

Compound table: links to PDB-related databases & PoSSuM

CofactorsSubstratesProductsintermediates
KEGG-idC00076C01330C00001C00012C00017C00012C00017I00087I00085I00086
CompoundCalciumSodiumH2OPeptideProteinPeptideProteinPeptidyl-tetrahedral intermediateAcyl-enzymeTetrahedral intermediate
Typedivalent metal (Ca2+, Mg2+)univalent metal (Na+, K+)H2Opeptide/proteinpeptide/proteinpeptide/proteinpeptide/protein


ChEBI29108
29101
15377







PubChem271
923
962
22247451







                  
1tecEBound:2x_CABound:_NA UnboundBound:LEU 45-ASP 46(chain I)UnboundUnbound   
2tecEBound:2x_CAUnbound UnboundBound:LEU 45-ASP 46(chain I)UnboundUnbound   
3tecEBound:3x_CAUnbound UnboundBound:LEU 45-ASP 46(chain I)UnboundUnbound   
1thmABound:2x_CABound:_NA UnboundUnboundUnboundUnbound   

Active-site residues
resource
Swiss-prot;P01051, P04072
pdbCatalytic residuesMain-chain involved in catalysis
          
1tecEASP 38;HIS 71;ASN 163;THR 224;SER 225
SER 225
2tecEASP 38;HIS 71;ASN 163;THR 224;SER 225
SER 225
3tecEASP 38;HIS 71;ASN 163;THR 224;SER 225
SER 225
1thmAASP 38;HIS 71;ASN 163;THR 224;SER 225
SER 225

References for Catalytic Mechanism
ReferencesSectionsNo. of steps in catalysis
[5]

[6]

[8]

[9]


references
[1]
PubMed ID6175064
JournalUltramicroscopy
Year1981
Volume7
Pages131-8
AuthorsSherman MB, Orlova EV, Terzyan SS, Kleine R, Kiselev NA
TitleOn the negative straining of the protein crystal structure.
[2]
PubMed ID3275467
JournalBiochim Biophys Acta
Year1988
Volume952
Pages20-6
AuthorsHausdorf G, Kruger K, Kuttner G, Holzhutter HG, Frommel C, Hohne WE
TitleOxidation of a methionine residue in subtilisin-type proteinases by the hydrogen peroxide/borate system--an active site-directed reaction.
[3]
PubMed ID3042463
JournalFEBS Lett
Year1988
Volume236
Pages171-8
AuthorsDauter Z, Betzel C, Hohne WE, Ingelman M, Wilson KS
TitleCrystal structure of a complex between thermitase from Thermoactinomyces vulgaris and the leech inhibitor eglin.
[4]
CommentsX-ray crystallography
PubMed ID2688688
JournalActa Crystallogr B
Year1989
Volume45
Pages488-99
AuthorsGros P, Fujinaga M, Dijkstra BW, Kalk KH, Hol WG
TitleCrystallographic refinement by incorporation of molecular dynamics: thermostable serine protease thermitase complexed with eglin c.
Related PDB1tec
[5]
CommentsX-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS).
Medline ID89171261
PubMed ID2647518
JournalFEBS Lett
Year1989
Volume244
Pages208-12
AuthorsTeplyakov AV, Kuranova IP, Harutyunyan EH, Frommel C, Hohne WE
TitleCrystal structure of thermitase from Thermoactinomyces vulgaris at 2.2 A resolution.
Related UniProtKBP04072
[6]
CommentsX-RAY CRYSTALLOGRAPHY (1.98 ANGSTROMS).
Medline ID90096158
PubMed ID2689655
JournalJ Mol Biol
Year1989
Volume210
Pages347-67
AuthorsGros P, Betzel C, Dauter Z, Wilson KS, Hol WG
TitleMolecular dynamics refinement of a thermitase-eglin-c complex at 1.98 A resolution and comparison of two crystal forms that differ in calcium content.
Related PDB2tec
Related UniProtKBP04072
[7]
PubMed ID2664764
JournalProteins
Year1989
Volume5
Pages22-37
AuthorsFrommel C, Sander C
TitleThermitase, a thermostable subtilisin: comparison of predicted and experimental structures and the molecular cause of thermostability.
[8]
CommentsX-RAY CRYSTALLOGRAPHY (1.4 ANGSTROMS).
Medline ID90317828
PubMed ID2196375
JournalJ Mol Biol
Year1990
Volume214
Pages261-79
AuthorsTeplyakov AV, Kuranova IP, Harutyunyan EH, Vainshtein BK, Frommel C, Hohne WE, Wilson KS
TitleCrystal structure of thermitase at 1.4 A resolution.
Related PDB1thm
Related UniProtKBP04072
[9]
PubMed ID2184432
JournalProtein Eng
Year1990
Volume3
Pages161-72
AuthorsBetzel C, Teplyakov AV, Harutyunyan EH, Saenger W, Wilson KS
TitleThermitase and proteinase K: a comparison of the refined three-dimensional structures of the native enzymes.
[10]
CommentsX-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS).
Medline ID91131592
PubMed ID1993669
JournalJ Biol Chem
Year1991
Volume266
Pages2953-61
AuthorsGros P, Kalk KH, Hol WG
TitleCalcium binding to thermitase. Crystallographic studies of thermitase at 0, 5, and 100 mM calcium.
Related PDB3tec
Related UniProtKBP04072
[11]
PubMed ID1553381
JournalProteins
Year1992
Volume12
Pages63-74
AuthorsGros P, Teplyakov AV, Hol WG
TitleEffects of eglin-c binding to thermitase: three-dimensional structure comparison of native thermitase and thermitase eglin-c complexes.
[12]
CommentsX-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS).
Medline ID93146156
PubMed ID8425603
JournalFEBS Lett
Year1993
Volume317
Pages185-8
AuthorsBetzel C, Dauter Z, Genov N, Lamzin V, Navaza J, Schnebli HP, Visanji M, Wilson KS
TitleStructure of the proteinase inhibitor eglin c with hydrolysed reactive centre at 2.0 A resolution.
Related UniProtKBP01051
[13]
PubMed ID8254666
JournalJ Mol Biol
Year1993
Volume234
Pages661-79
AuthorsKrystek S, Stouch T, Novotny J
TitleAffinity and specificity of serine endopeptidase-protein inhibitor interactions. Empirical free energy calculations based on X-ray crystallographic structures.
[14]
PubMed ID7558600
JournalInt J Pept Protein Res
Year1995
Volume46
Pages73-8
AuthorsBrandt W, Lehmann T, Willkomm C, Fittkau S, Barth A
TitleCoMFA investigations on two series of artificial peptide inhibitors of the serine protease thermitase. Synthesis of an inhibitor of predicted greater potency.
[15]
PubMed ID8796317
JournalAdv Exp Med Biol
Year1996
Volume379
Pages133-40
AuthorsPeters K, Bromme D, Jahreis G, Fittkau S
TitleThermitase - kinetic differentiation to the subtilisins.
[16]
PubMed ID8796305
JournalAdv Exp Med Biol
Year1996
Volume379
Pages5-9
AuthorsTeplyakov A, Gros P, Hol WG
TitleCrystallographic study of eglin-C binding to thermitase.
[17]
PubMed ID9048543
JournalBiochemistry
Year1997
Volume36
Pages1598-607
AuthorsQasim MA, Ganz PJ, Saunders CW, Bateman KS, James MN, Laskowski M Jr
TitleInterscaffolding additivity. Association of P1 variants of eglin c and of turkey ovomucoid third domain with serine proteinases.
[18]
PubMed ID9605545
JournalProtein Eng
Year1998
Volume11
Pages109-17
AuthorsMei HC, Liaw YC, Li YC, Wang DC, Takagi H, Tsai YC
TitleEngineering subtilisin YaB: restriction of substrate specificity by the substitution of Gly124 and Gly151 with Ala.

comments
This enzyme belongs to the peptidase family-S8.
The calcium ions and sodium ion play a structural role, rather than a catalytic one.
This enzyme contains a classic catalytic triad (Ser/His/Asp), suggesting that it must have a similar mechanism to that of trypsin (D00197 in EzCatDB). However, in contrast to trypsin-like enzymes (where mainchain amide groups form an oxyanion hole), sidechains of Asn163 and Thr224 may form an oxyanion hole, which stabilizes the transition-state, together with the mainchain amide group of Ser225, as in subtilisin (D00219 in EzCatDB).

createdupdated
2004-08-242011-02-21


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