DB code: S00519

RLCP classification 1.13.30000.9 : Hydrolysis
CATH domain 3.40.50.200 : Rossmann fold Catalytic domain
E.C. 3.4.21.66
CSA
M-CSA
MACiE

CATH domain Related DB codes (homologues)
3.40.50.200 : Rossmann fold S00295 S00296 D00219

Uniprot Enzyme Name
UniprotKB Protein name Synonyms MEROPS Pfam
P04072 Thermitase
EC 3.4.21.66
S08.007 (Serine)
PF00082 (Peptidase_S8)
[Graphical View]

KEGG enzyme name
thermitase
thermophilic Streptomyces serine proteinase
Thermoactinomyces vulgaris serine proteinase

UniprotKB: Accession Number Entry name Activity Subunit Subcellular location Cofactor
P04072 THET_THEVU Hydrolysis of proteins, including collagen. Secreted. Binds 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.

KEGG Pathways
Map code Pathways E.C.

Compound table
Cofactors Substrates Products Intermediates
KEGG-id C00076 C01330 C00001 C00012 C00017 C00012 C00017 I00087 I00085 I00086
E.C.
Compound Calcium Sodium H2O Peptide Protein Peptide Protein Peptidyl-tetrahedral intermediate Acyl-enzyme Tetrahedral intermediate
Type divalent metal (Ca2+, Mg2+) univalent metal (Na+, K+) H2O peptide/protein peptide/protein peptide/protein peptide/protein
ChEBI 29108
29101
15377
PubChem 271
923
22247451
962
1tecE Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Bound:2x_CA Bound:_NA Unbound Bound:LEU_45-ASP_46(chain I) Unbound Unbound
2tecE Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Bound:2x_CA Unbound Unbound Bound:LEU_45-ASP_46(chain I) Unbound Unbound
3tecE Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Bound:3x_CA Unbound Unbound Bound:LEU_45-ASP_46(chain I) Unbound Unbound
1thmA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Bound:2x_CA Bound:_NA Unbound Unbound Unbound Unbound

Reference for Active-site residues
resource references E.C.
Swiss-prot;P01051, P04072

Active-site residues
PDB Catalytic residues Cofactor-binding residues Modified residues Main-chain involved in catalysis Comment
1tecE Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASP 38;HIS 71;ASN 163;THR 224;SER 225 SER 225
2tecE Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASP 38;HIS 71;ASN 163;THR 224;SER 225 SER 225
3tecE Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASP 38;HIS 71;ASN 163;THR 224;SER 225 SER 225
1thmA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASP 38;HIS 71;ASN 163;THR 224;SER 225 SER 225

References for Catalytic Mechanism
References Sections No. of steps in catalysis
[5]
[6]
[8]
[9]

References
[1]
Resource
Comments
Medline ID
PubMed ID 6175064
Journal Ultramicroscopy
Year 1981
Volume 7
Pages 131-8
Authors Sherman MB, Orlova EV, Terzyan SS, Kleine R, Kiselev NA
Title On the negative straining of the protein crystal structure.
Related PDB
Related UniProtKB
[2]
Resource
Comments
Medline ID
PubMed ID 3275467
Journal Biochim Biophys Acta
Year 1988
Volume 952
Pages 20-6
Authors Hausdorf G, Kruger K, Kuttner G, Holzhutter HG, Frommel C, Hohne WE
Title Oxidation of a methionine residue in subtilisin-type proteinases by the hydrogen peroxide/borate system--an active site-directed reaction.
Related PDB
Related UniProtKB
[3]
Resource
Comments
Medline ID
PubMed ID 3042463
Journal FEBS Lett
Year 1988
Volume 236
Pages 171-8
Authors Dauter Z, Betzel C, Hohne WE, Ingelman M, Wilson KS
Title Crystal structure of a complex between thermitase from Thermoactinomyces vulgaris and the leech inhibitor eglin.
Related PDB
Related UniProtKB
[4]
Resource
Comments X-ray crystallography
Medline ID
PubMed ID 2688688
Journal Acta Crystallogr B
Year 1989
Volume 45
Pages 488-99
Authors Gros P, Fujinaga M, Dijkstra BW, Kalk KH, Hol WG
Title Crystallographic refinement by incorporation of molecular dynamics: thermostable serine protease thermitase complexed with eglin c.
Related PDB 1tec
Related UniProtKB
[5]
Resource
Comments X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS).
Medline ID 89171261
PubMed ID 2647518
Journal FEBS Lett
Year 1989
Volume 244
Pages 208-12
Authors Teplyakov AV, Kuranova IP, Harutyunyan EH, Frommel C, Hohne WE
Title Crystal structure of thermitase from Thermoactinomyces vulgaris at 2.2 A resolution.
Related PDB
Related UniProtKB P04072
[6]
Resource
Comments X-RAY CRYSTALLOGRAPHY (1.98 ANGSTROMS).
Medline ID 90096158
PubMed ID 2689655
Journal J Mol Biol
Year 1989
Volume 210
Pages 347-67
Authors Gros P, Betzel C, Dauter Z, Wilson KS, Hol WG
Title Molecular 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 PDB 2tec
Related UniProtKB P04072
[7]
Resource
Comments
Medline ID
PubMed ID 2664764
Journal Proteins
Year 1989
Volume 5
Pages 22-37
Authors Frommel C, Sander C
Title Thermitase, a thermostable subtilisin: comparison of predicted and experimental structures and the molecular cause of thermostability.
Related PDB
Related UniProtKB
[8]
Resource
Comments X-RAY CRYSTALLOGRAPHY (1.4 ANGSTROMS).
Medline ID 90317828
PubMed ID 2196375
Journal J Mol Biol
Year 1990
Volume 214
Pages 261-79
Authors Teplyakov AV, Kuranova IP, Harutyunyan EH, Vainshtein BK, Frommel C, Hohne WE, Wilson KS
Title Crystal structure of thermitase at 1.4 A resolution.
Related PDB 1thm
Related UniProtKB P04072
[9]
Resource
Comments
Medline ID
PubMed ID 2184432
Journal Protein Eng
Year 1990
Volume 3
Pages 161-72
Authors Betzel C, Teplyakov AV, Harutyunyan EH, Saenger W, Wilson KS
Title Thermitase and proteinase K: a comparison of the refined three-dimensional structures of the native enzymes.
Related PDB
Related UniProtKB
[10]
Resource
Comments X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS).
Medline ID 91131592
PubMed ID 1993669
Journal J Biol Chem
Year 1991
Volume 266
Pages 2953-61
Authors Gros P, Kalk KH, Hol WG
Title Calcium binding to thermitase. Crystallographic studies of thermitase at 0, 5, and 100 mM calcium.
Related PDB 3tec
Related UniProtKB P04072
[11]
Resource
Comments
Medline ID
PubMed ID 1553381
Journal Proteins
Year 1992
Volume 12
Pages 63-74
Authors Gros P, Teplyakov AV, Hol WG
Title Effects of eglin-c binding to thermitase: three-dimensional structure comparison of native thermitase and thermitase eglin-c complexes.
Related PDB
Related UniProtKB
[12]
Resource
Comments X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS).
Medline ID 93146156
PubMed ID 8425603
Journal FEBS Lett
Year 1993
Volume 317
Pages 185-8
Authors Betzel C, Dauter Z, Genov N, Lamzin V, Navaza J, Schnebli HP, Visanji M, Wilson KS
Title Structure of the proteinase inhibitor eglin c with hydrolysed reactive centre at 2.0 A resolution.
Related PDB
Related UniProtKB P01051
[13]
Resource
Comments
Medline ID
PubMed ID 8254666
Journal J Mol Biol
Year 1993
Volume 234
Pages 661-79
Authors Krystek S, Stouch T, Novotny J
Title Affinity and specificity of serine endopeptidase-protein inhibitor interactions. Empirical free energy calculations based on X-ray crystallographic structures.
Related PDB
Related UniProtKB
[14]
Resource
Comments
Medline ID
PubMed ID 7558600
Journal Int J Pept Protein Res
Year 1995
Volume 46
Pages 73-8
Authors Brandt W, Lehmann T, Willkomm C, Fittkau S, Barth A
Title CoMFA investigations on two series of artificial peptide inhibitors of the serine protease thermitase. Synthesis of an inhibitor of predicted greater potency.
Related PDB
Related UniProtKB
[15]
Resource
Comments
Medline ID
PubMed ID 8796317
Journal Adv Exp Med Biol
Year 1996
Volume 379
Pages 133-40
Authors Peters K, Bromme D, Jahreis G, Fittkau S
Title Thermitase - kinetic differentiation to the subtilisins.
Related PDB
Related UniProtKB
[16]
Resource
Comments
Medline ID
PubMed ID 8796305
Journal Adv Exp Med Biol
Year 1996
Volume 379
Pages 5-9
Authors Teplyakov A, Gros P, Hol WG
Title Crystallographic study of eglin-C binding to thermitase.
Related PDB
Related UniProtKB
[17]
Resource
Comments
Medline ID
PubMed ID 9048543
Journal Biochemistry
Year 1997
Volume 36
Pages 1598-607
Authors Qasim MA, Ganz PJ, Saunders CW, Bateman KS, James MN, Laskowski M Jr
Title Interscaffolding additivity. Association of P1 variants of eglin c and of turkey ovomucoid third domain with serine proteinases.
Related PDB
Related UniProtKB
[18]
Resource
Comments
Medline ID
PubMed ID 9605545
Journal Protein Eng
Year 1998
Volume 11
Pages 109-17
Authors Mei HC, Liaw YC, Li YC, Wang DC, Takagi H, Tsai YC
Title Engineering subtilisin YaB: restriction of substrate specificity by the substitution of Gly124 and Gly151 with Ala.
Related PDB
Related UniProtKB

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).

Created Updated
2004-08-24 2011-02-21