DB code: D00442

RLCP classification 1.13.200.966 : Hydrolysis
CATH domain 2.40.70.10 : Cathepsin D, subunit A; domain 1 Catalytic 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
PubChem 22247451
962
1mppA01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound
2asiA01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound
2rmpA01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Analogue:IVA-VAL-VAL-STA-ALA-STA (chain B) Unbound Unbound Unbound Unbound Unbound
1mppA02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound Unbound
2asiA02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Bound:HOH_421 Unbound Unbound Unbound
2rmpA02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain 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 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASP 32
2asiA01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASP 38
2rmpA01 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASP 38
1mppA02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASP 215
2asiA02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASP 237
2rmpA02 Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain 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.

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