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 29108	29101 29101	15377 15377
PubChem	271 271	923 923	22247451 962 22247451 962

1tecE	Bound:2x_CA	Bound:_NA		Unbound	Bound:LEU_45-ASP_46(chain I)	Unbound	Unbound
2tecE	Bound:2x_CA	Unbound		Unbound	Bound:LEU_45-ASP_46(chain I)	Unbound	Unbound
3tecE	Bound:3x_CA	Unbound		Unbound	Bound:LEU_45-ASP_46(chain I)	Unbound	Unbound
1thmA	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	ASP 38;HIS 71;ASN 163;THR 224;SER 225			SER 225
2tecE	ASP 38;HIS 71;ASN 163;THR 224;SER 225			SER 225
3tecE	ASP 38;HIS 71;ASN 163;THR 224;SER 225			SER 225
1thmA	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).

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