DB code: S00296

RLCP classification	1.13.30000.12 : Hydrolysis
CATH domain	3.40.50.200 : Rossmann fold	Catalytic domain
E.C.	3.4.21.100
CSA
M-CSA
MACiE

CATH domain	Related DB codes (homologues)
3.40.50.200 : Rossmann fold	S00295 D00219 S00519

Uniprot Enzyme Name
UniprotKB	Protein name	Synonyms	MEROPS	Pfam
P42790	Pseudomonalisin	EC 3.4.21.100 Pepstatin-insensitive carboxyl proteinase Pseudomonapepsin	S53.001 (Serine)	PF00082 (Peptidase_S8) PF09286 (Pro-kuma_activ) [Graphical View]

KEGG enzyme name
sedolisin Pseudomonas sp. pepstatin-insensitive carboxyl proteinase pseudomonapepsin pseudomonalisin sedolysin

UniprotKB: Accession Number	Entry name	Activity	Subunit	Subcellular location	Cofactor
P42790	PICP_PSESR	Hydrolysis of the B chain of insulin at 13- Glu-\|-Ala-14, 15-Leu-\|-Tyr-16 and 25-Phe-\|-Tyr-26 and angiotensin I at 4-Tyr-\|-Ile-5. A good synthetic substrate is Lys-Pro-Ile-Glu- Phe-\|-Phe(NO(2))-Arg-Leu.		Periplasm.	Binds 1 calcium ion per subunit.

KEGG Pathways
Map code	Pathways	E.C.

Compound table
	Substrates		Products	Intermediates
KEGG-id	C00017	C00001	C00012	I00087	I00085	I00086
E.C.
Compound	Protein	H2O	Peptide	Peptidyl-tetrahedral intermediate	Acyl-enzyme	Tetrahedral intermediate
Type	peptide/protein	H2O	peptide/protein
ChEBI		15377 15377
PubChem		22247451 962 22247451 962

1ga1A	Unbound		Bound:UNK-PHI-UNK	Unbound	Unbound	Unbound
1ga4A	Unbound		Unbound	Unbound	Intermediate-bound:IVA-PHI-TYB	Unbound
1ga6A	Unbound		Unbound	Unbound	Unbound	Unbound
1kdvA	Unbound		Unbound	Unbound	Intermediate-bound:ACE-ILE-ALA-PHA	Unbound
1kdyA	Unbound		Unbound	Unbound	Intermediate-bound:ACE-ILE-PRO-PHA	Unbound
1kdzA	Unbound		Unbound	Unbound	Unbound	Transition-state-analogue:IVA-TYR-LEU-TYB
1ke1A	Unbound		Unbound	Unbound	Unbound	Transition-state-analogue:IVA-TYR-TYB
1ke2A	Unbound		Unbound	Unbound	Unbound	Transition-state-analogue:CSI-LEU-PHA

Reference for Active-site residues
resource	references	E.C.
Swiss-prot & literature [1],[3]

Active-site residues
PDB	Catalytic residues	Cofactor-binding residues	Modified residues	Main-chain involved in catalysis	Comment

1ga1A	GLU 80;ASP 84;ASP 170;SER 287			SER 287
1ga4A	GLU 80;ASP 84;ASP 170;SER 287			SER 287
1ga6A	GLU 80;ASP 84;ASP 170;SER 287			SER 287
1kdvA	GLU 80;ASP 84;ASP 170;SER 287			SER 287
1kdyA	GLU 80;ASP 84;ASP 170;SER 287			SER 287
1kdzA	GLU 80;ASP 84;ASP 170;SER 287			SER 287
1ke1A	GLU 80;ASP 84;ASP 170;SER 287			SER 287
1ke2A	GLU 80;ASP 84;ASP 170;SER 287			SER 287

References for Catalytic Mechanism
References	Sections	No. of steps in catalysis
[3]	p.445-446
[4]	p.15609
[5]	p.88-92

References
[1]
Resource
Comments	active site (mutation analysis)
Medline ID	99419069
PubMed ID	10488127
Journal	J Biol Chem
Year	1999
Volume	274
Pages	27815-22
Authors	Hiroshi Oyama, Shin-ichiro Abe, Souko Ushiyama, Saori Takahashi, and Kohei Oda
Title	Identification of Catalytic Residues of Pepstatin-insensitive Carboxyl Proteinases from Prokaryotes by Site-directed Mutagenesis
Related PDB	1ga6
Related UniProtKB	P42790
[2]
Resource
Comments	X-ray crystallography
Medline ID
PubMed ID	11173470
Journal	Acta Crystallogr D
Year	2001
Volume	57
Pages	239-49
Authors	Dauter Z, Li M, Wlodawer A
Title	Practical experience with the use of halides for phasing macromolecular structures: a powerful tool for structural genomics.
Related PDB	1ga1
Related UniProtKB
[3]
Resource
Comments	X-ray crystallography (1.0-1.4 Angstroms)
Medline ID
PubMed ID	11323721
Journal	Nat Struct Biol
Year	2001
Volume	8
Pages	442-6
Authors	Wlodawer A, Li M, Dauter Z, Gustchina A, Uchida K, Oyama H, Dunn BM, Oda K
Title	Carboxyl proteinase from Pseudomonas defines a novel family of subtilisin-like enzymes.
Related PDB	1ga4 1ga6
Related UniProtKB
[4]
Resource
Comments
Medline ID
PubMed ID	11747435
Journal	Biochemistry
Year	2001
Volume	40
Pages	15602-11
Authors	Wlodawer A, Li M, Gustchina A, Dauter Z, Uchida K, Oyama H, Goldfarb NE, Dunn BM, Oda K
Title	Inhibitor complexes of the Pseudomonas serine-carboxyl proteinase.
Related PDB	1kdv 1kdy 1kdz 1ke1 1ke2
Related UniProtKB
[5]
Resource
Comments	Review
Medline ID
PubMed ID	12673349
Journal	Acta Biochim Pol
Year	2003
Volume	50
Pages	81-102
Authors	Wlodawer A, Li M, Gustchina A, Oyama H, Dunn BM, Oda K
Title	Structural and enzymatic properties of the sedolisin family of serine-carboxyl peptidases.
Related PDB
Related UniProtKB

Comments
According to the papers [3], [4] & [5], Ser287 and the acidic residues seem to comprise the catalytic residues, although the detailed catalytic mechanism remains still unknown. However, the catalytic serine residue plays a role as nucleophile, which will attack the carbonyl carbon atom and make a covalent bond with the atom (see [3], [4] & [5]). The paper [3] suggested that Glu80 might be a general base that will abstract the proton of the -OH group of the catalytic serine. Thus, Ser287-Glu80-Asp84 constitute the catalytic triad, whilst Asp170 seems to be a part of oxyanion hole stabilizing the tetrahedral intermediate of the reaction (see [3] & [4]).

Comments

According to the papers [3], [4] & [5], Ser287 and the acidic residues seem to comprise the catalytic residues, although the detailed catalytic mechanism remains still unknown. However, the catalytic serine residue plays a role as nucleophile, which will attack the carbonyl carbon atom and make a covalent bond with the atom (see [3], [4] & [5]). The paper [3] suggested that Glu80 might be a general base that will abstract the proton of the -OH group of the catalytic serine. Thus, Ser287-Glu80-Asp84 constitute the catalytic triad, whilst Asp170 seems to be a part of oxyanion hole stabilizing the tetrahedral intermediate of the reaction (see [3] & [4]).

Created	Updated
2002-07-04	2011-02-22