DB code: D00442

RLCP classification	1.13.200.966 : Hydrolysis
CATH domain	2.40.70.10 : Cathepsin D, subunit A; domain 1	Catalytic domain
CATH domain	2.40.70.10 : Cathepsin D, subunit A; domain 1	Catalytic domain
E.C.	3.4.23.23
CSA	1mpp
M-CSA	1mpp
MACiE

CATH domain	Related DB codes (homologues)
2.40.70.10 : Cathepsin D, subunit A; domain 1	D00471 D00436 D00438 D00439 D00440 D00441 D00443 D00437 D00444 D00423 D00445 D00484 M00206 M00166 D00231 D00529

Uniprot Enzyme Name
UniprotKB	Protein name	Synonyms	MEROPS	Pfam
P00799	Mucorpepsin	EC 3.4.23.23 Mucor rennin	A01.013 (Aspartic)	PF00026 (Asp) [Graphical View]
P09177	Mucorpepsin	EC 3.4.23.23 Mucor rennin	A01.013 (Aspartic)	PF00026 (Asp) [Graphical View]

KEGG enzyme name
mucorpepsin Mucor rennin Mucor aspartic proteinase Mucor acid proteinase Mucor acid protease Mucor miehei aspartic proteinase Mucor miehei aspartic protease Mucor aspartic proteinase Mucor pusillus emporase Fromase 100 Mucor pusillus rennin Fromase 46TL Mucor miehei rennin

UniprotKB: Accession Number	Entry name	Activity	Subunit	Subcellular location	Cofactor
P00799	CARP_RHIMI	Hydrolysis of proteins, favoring hydrophobic residues at P1 and P1''. Clots milk. Does not accept Lys at P1, and hence does not activate trypsinogen.
P09177	CARP_RHIPU	Hydrolysis of proteins, favoring hydrophobic residues at P1 and P1''. Clots milk. Does not accept Lys at P1, and hence does not activate trypsinogen.

KEGG Pathways
Map code	Pathways	E.C.

Compound table
	Substrates				Products		Intermediates
KEGG-id	C00017	C00012	L00078	C00001	C00017	C00012	I00136
E.C.
Compound	Protein	Peptide	Casein	H2O	Protein	Peptide	Amino-diol-tetrahedral intermediate
Type	peptide/protein	peptide/protein	peptide/protein,phosphate group/phosphate ion	H2O	peptide/protein	peptide/protein
ChEBI				15377 15377
PubChem				22247451 962 22247451 962

1mppA01	Unbound	Unbound	Unbound		Unbound	Unbound	Unbound
2asiA01	Unbound	Unbound	Unbound		Unbound	Unbound	Unbound
2rmpA01	Analogue:IVA-VAL-VAL-STA-ALA-STA (chain B)	Unbound	Unbound		Unbound	Unbound	Unbound
1mppA02	Unbound	Unbound	Unbound		Unbound	Unbound	Unbound
2asiA02	Unbound	Unbound	Unbound	Bound:HOH_421	Unbound	Unbound	Unbound
2rmpA02	Unbound	Unbound	Unbound		Unbound	Unbound	Unbound

Reference for Active-site residues
resource	references	E.C.
PDB;1e5o,1e80,1e81,1e82,1mpp,2asi & Swiss-prot;P00799,P09177

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

1mppA01	ASP 32
2asiA01	ASP 38
2rmpA01	ASP 38
1mppA02	ASP 215
2asiA02	ASP 237
2rmpA02	ASP 237

References for Catalytic Mechanism
References	Sections	No. of steps in catalysis
[3]	Fig.5, p.7010-7012	3
[12]	p.453-456

References
[1]
Resource
Comments
Medline ID
PubMed ID	1125813
Journal	Can J Biochem
Year	1975
Volume	53
Pages	269-74
Authors	Rickert WS, McBride-Warren PA
Title	Acid proteases from species of Mucormii. partial characterization of the acid protease produced by a strain of Mucor miehei isolated in Cuba.
Related PDB
Related UniProtKB
[2]
Resource
Comments
Medline ID
PubMed ID	6749830
Journal	J Biochem (Tokyo)
Year	1982
Volume	91
Pages	2039-46
Authors	Etoh Y, Shoun H, Ogino T, Fujiwara S, Arima K, Beppu T
Title	Proton magnetic resonance spectroscopy of an essential histidyl residue in a milk-clotting acid protease, Mucor rennin.
Related PDB
Related UniProtKB
[3]
Resource
Comments
Medline ID
PubMed ID	3313384
Journal	Proc Natl Acad Sci U S A
Year	1987
Volume	84
Pages	7009-13
Authors	Suguna K, Padlan EA, Smith CW, Carlson WD, Davies DR
Title	Binding of a reduced peptide inhibitor to the aspartic proteinase from Rhizopus chinensis: implications for a mechanism of action.
Related PDB
Related UniProtKB
[4]
Resource
Comments
Medline ID
PubMed ID	1963762
Journal	Ann N Y Acad Sci
Year	1990
Volume	613
Pages	14-25
Authors	Beppu T
Title	Modification of milk-clotting aspartic proteinases by recombinant DNA techniques.
Related PDB
Related UniProtKB
[5]
Resource
Comments
Medline ID
PubMed ID	2116411
Journal	J Biol Chem
Year	1990
Volume	265
Pages	13955-9
Authors	Aikawa J, Yamashita T, Nishiyama M, Horinouchi S, Beppu T
Title	Effects of glycosylation on the secretion and enzyme activity of Mucor rennin, an aspartic proteinase of Mucor pusillus, produced by recombinant yeast.
Related PDB
Related UniProtKB
[6]
Resource
Comments
Medline ID
PubMed ID	1892395
Journal	Appl Environ Microbiol
Year	1991
Volume	57
Pages	2052-6
Authors	Hiramatsu R, Horinouchi S, Uchida E, Hayakawa T, Beppu T
Title	The secretion leader of Mucor pusillus rennin which possesses an artificial Lys-Arg sequence directs the secretion of mature human growth hormone by Saccharomyces cerevisiae.
Related PDB
Related UniProtKB
[7]
Resource
Comments
Medline ID
PubMed ID	2001389
Journal	Biochim Biophys Acta
Year	1991
Volume	1076
Pages	406-15
Authors	Brown ED, Yada RY
Title	A kinetic and equilibrium study of the denaturation of aspartic proteinases from the fungi, Endothia parasitica and Mucor miehei.
Related PDB
Related UniProtKB
[8]
Resource
Comments	X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS).
Medline ID	93195933
PubMed ID	8450540
Journal	J Mol Biol
Year	1993
Volume	230
Pages	260-83
Authors	Newman M, Watson F, Roychowdhury P, Jones H, Badasso M, Cleasby A, Wood SP, Tickle IJ, Blundell TL
Title	X-ray analyses of aspartic proteinases. V. Structure and refinement at 2.0 A resolution of the aspartic proteinase from Mucor pusillus.
Related PDB	1mpp
Related UniProtKB	P09177
[9]
Resource
Comments	X-RAY CRYSTALLOGRAPHY (2.8 ANGSTROMS).
Medline ID
PubMed ID
Journal	Acta Crystallogr D Biol Crystallogr
Year	1995
Volume	51
Pages	243-4
Authors	Jia Z, Vandonselaar M, Schneider P, Quail JW
Title	Crystallization and preliminary X-ray structure solution of Rhizomucor miehei aspartic proteinase
Related PDB
Related UniProtKB	P00799
[10]
Resource
Comments
Medline ID
PubMed ID	8540364
Journal	Adv Exp Med Biol
Year	1995
Volume	362
Pages	501-9
Authors	Beppu T, Park YN, Aikawa J, Nishiyama M, Horinouchi S
Title	Tyrosine 75 on the flap contributes to enhance catalytic efficiency of a fungal aspartic proteinase, Mucor pusillus pepsin.
Related PDB
Related UniProtKB
[11]
Resource
Comments
Medline ID
PubMed ID	9058201
Journal	J Biochem (Tokyo)
Year	1997
Volume	121
Pages	118-21
Authors	Park YN, Aikawa J, Nishiyama M, Horinouchi S, Beppu T
Title	Site-directed mutagenesis of conserved Trp39 in Rhizomucor pusillus pepsin: possible role of Trp39 in maintaining Tyr75 in the correct orientation for maximizing catalytic activity.
Related PDB
Related UniProtKB
[12]
Resource
Comments	X-RAY CRYSTALLOGRAPHY (2.15 ANGSTROMS).
Medline ID	97303051
PubMed ID	9159482
Journal	J Mol Biol
Year	1997
Volume	268
Pages	449-59
Authors	Yang J, Teplyakov A, Quail JW
Title	Crystal structure of the aspartic proteinase from Rhizomucor miehei at 2.15 A resolution.
Related PDB	2asi
Related UniProtKB	P00799
[13]
Resource
Comments
Medline ID
PubMed ID	9561231
Journal	Adv Exp Med Biol
Year	1998
Volume	436
Pages	283-92
Authors	Quail JW, Yang J, Schneider P, Jia Z
Title	Crystal structure of the Rhizomucor miehei aspartic proteinase.
Related PDB
Related UniProtKB
[14]
Resource
Comments	X-ray crystallography
Medline ID
PubMed ID	10089458
Journal	Acta Crystallogr D Biol Crystallogr
Year	1999
Volume	55
Pages	625-30
Authors	Yang J, Quail JW
Title	Structure of the Rhizomucor miehei aspartic proteinase complexed with the inhibitor pepstatin A at 2.7 A resolution.
Related PDB	2rmp
Related UniProtKB
[15]
Resource
Comments
Medline ID
PubMed ID	10744950
Journal	Biotechnol Appl Biochem
Year	2000
Volume	31
Pages	77-84
Authors	Beldarrain A, Acosta N, Montesinos R, Mata M, Cremata J
Title	Characterization of Mucor pusillus rennin expressed in Pichia pastoris: enzymic, spectroscopic and calorimetric studies.
Related PDB
Related UniProtKB
[16]
Resource
Comments
Medline ID
PubMed ID	11328603
Journal	J Biochem (Tokyo)
Year	2001
Volume	129
Pages	791-4
Authors	Aikawa J, Park YN, Sugiyama M, Nishiyama M, Horinouchi S, Beppu T
Title	Replacements of amino acid residues at subsites and their effects on the catalytic properties of Rhizomucor pusillus pepsin, an aspartic proteinase from Rhizomucor pusillus.
Related PDB
Related UniProtKB

Comments
This enzyme belongs to the peptidae family-A1. According to the literature [12], this enzyme has got a catalytic dyad composed of two aspartate residues, supporting the catalytic mechanism proposed by the paper [3]. Accoriding to the proposed mechanism (see [3]), the sidechains of both the aspartic acid residues are hydrogen-bonded to the catalytic water. The sidechain of the ionized aspartate (possibly corresponding to Asp215 of 1mpp) might act as a general base, which can abstract a proton from the water, which in turn would make a nucleophilic attack on the carbonyl carbon of the peptide bond. Meanwhile, the protonated sidechain of the other aspartate (corresponding to Asp32 of 1mpp) may stabilize the negative charge on the carbonyl oxygen of the scissile bond during the transition state. At the next stage, the sidechain of the aspartate that had accepted a proton from water could protonate the leaving nitrogen atom, as a general acid, during the cleavage of the peptide bond.

Comments

This enzyme belongs to the peptidae family-A1.
According to the literature [12], this enzyme has got a catalytic dyad composed of two aspartate residues, supporting the catalytic mechanism proposed by the paper [3].
Accoriding to the proposed mechanism (see [3]), the sidechains of both the aspartic acid residues are hydrogen-bonded to the catalytic water.
The sidechain of the ionized aspartate (possibly corresponding to Asp215 of 1mpp) might act as a general base, which can abstract a proton from the water, which in turn would make a nucleophilic attack on the carbonyl carbon of the peptide bond.
Meanwhile, the protonated sidechain of the other aspartate (corresponding to Asp32 of 1mpp) may stabilize the negative charge on the carbonyl oxygen of the scissile bond during the transition state.
At the next stage, the sidechain of the aspartate that had accepted a proton from water could protonate the leaving nitrogen atom, as a general acid, during the cleavage of the peptide bond.

Created	Updated
2004-04-30	2012-06-28