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Information on EC 3.4.22.51 - cruzipain

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EC Tree
     3 Hydrolases
         3.4 Acting on peptide bonds (peptidases)
             3.4.22 Cysteine endopeptidases
                3.4.22.51 cruzipain
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This record set is specific for:
UNIPROT: Q598P5 not found.
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Word Map
The enzyme appears in viruses and cellular organisms
Reaction Schemes
broad endopeptidase specificity similar to that of cathepsin L
Synonyms
cruzipain, cruzain, rhodesain, major cysteine proteinase, congopain, cathepsin l-like cysteine protease, cathepsin l-like protease, gp57/51, brucipain, trypanosoma cruzi cysteine protease, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
congopain
-
-
-
-
cruzain
-
-
-
-
evansain
-
-
-
-
GP57/51
-
-
-
-
proteinase, Trypanosoma congolese cysteine
-
-
-
-
proteinase, Trypanosoma cruzi cysteine
-
-
-
-
proteinase, Trypanosoma cysteine
-
-
-
-
trypanopain
-
-
-
-
Trypanosoma congolese cysteine protease
-
-
-
-
Trypanosoma cruzi cysteine protease
-
-
-
-
Trypanosoma cysteine protease
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY hide
102227-51-0
-
141588-22-9
-
84399-99-5
-
90371-53-2
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2-aminobenzoic acid-ALRFSKQ-N-(2,4-dinitrophenyl)ethylenediamine + H2O
?
show the reaction diagram
-
-
-
?
2-aminobenzoic acid-KLGFSKQ-N-(2,4-dinitrophenyl)ethylenediamine + H2O
?
show the reaction diagram
-
-
-
?
2-aminobenzoic acid-KLRFSKQ-N-(2,4-dinitrophenyl)ethylenediamine + H2O
?
show the reaction diagram
and analogues
-
-
?
2-aminobenzoic acid-peptidyl-N-(2,4-dinitrophenyl)ethylenediamine + H2O
?
show the reaction diagram
and derivatives, enzyme shows preference for benzyl-Cys or Arg at the P1 position and a hydrophobic non-aromatic residue at position P2
-
-
?
2-aminobenzoyl-LGMISLMKRPQ-N-(2,4-dinitrophenyl)ethylenediamine + H2O
?
show the reaction diagram
azocasein + H2O
?
show the reaction diagram
benzoyl-FR-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
benzoyl-His-Arg-7-amido-4-methylcoumarin + H2O
benzoyl-His-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
benzoyl-HR-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
less susceptible substrate
-
-
?
benzoyl-PFR-4-nitroanilide + H2O
?
show the reaction diagram
benzoyl-Phe-Arg-4-methylcoumarin-7-amide + H2O
?
show the reaction diagram
fluorogenic substrates based on the peptide benzoyl-Phe-Arg-4-methylcoumarin-7-amide
-
-
?
benzoyl-Phe-Arg-7-amido-4-methylcoumarin + H2O
benzoyl-Phe-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
benzoyl-RR-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
less susceptible substrate
-
-
?
benzoyl-Tyr-Arg-7-amido-4-methylcoumarin + H2O
benzoyl-Tyr-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
-
-
-
?
benzoyl-YR-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
-
-
-
?
benzyloxycarbonyl-Phe-Arg-4-methylcoumarin 7-amide + H2O
?
show the reaction diagram
Bovine serum albumin + H2O
?
show the reaction diagram
-
-
-
?
casein + H2O
?
show the reaction diagram
-
-
-
?
Hemoglobin + H2O
?
show the reaction diagram
-
-
-
?
Immunoglobulin G1 + H2O
?
show the reaction diagram
Immunoglobulin G3 + H2O
?
show the reaction diagram
insulin A + H2O
?
show the reaction diagram
-
-
-
?
insulin B + H2O
?
show the reaction diagram
-
-
-
?
KEEASSAVVGGPG + H2O
?
show the reaction diagram
KEEASSAVVRGPG + H2O
?
show the reaction diagram
N-carbobenzoxy-Arg-Arg-2-(4-methoxy)-naphthylamide + H2O
N-carbobenzoxy-Arg-Arg + 2-amino-4-methoxynaphthalene
show the reaction diagram
-
-
-
?
N-carbobenzoxy-Val-Val-Arg-4-nitroanilide + H2O
?
show the reaction diagram
N-carbobenzoxy-VVR-4-nitroanilide + H2O
N-carbobenzoxy-VVR + 4-nitroaniline
show the reaction diagram
peptidyl-4-nitroanilide + H2O
?
show the reaction diagram
derivatives of
-
-
?
peptidyl-beta-naphthylamide + H2O
?
show the reaction diagram
peptidyl-methylcoumarin amide + H2O
?
show the reaction diagram
and derivatives, enzyme shows preference for benzyl-Cys or Arg at the P1 position and a hydrophobic non-aromatic residue at position P2
-
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
casein + H2O
?
show the reaction diagram
-
-
-
?
Hemoglobin + H2O
?
show the reaction diagram
-
-
-
?
Immunoglobulin G1 + H2O
?
show the reaction diagram
-
-
-
?
Immunoglobulin G3 + H2O
?
show the reaction diagram
-
-
-
?
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
NaCl
Km increases up to 400 mM, then decreases and remains constant at 50-60% up to 1 M, kcat maximum at 300 mM
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
alpha2-Macroglobulin
40% inhibition at a 1:1 ratio of cruzipain and alpha2-macroglobulin
-
azocasein
substrate inhibition
-
benzyloxycarbonyl-Phe-Ala-fluoromethylketone
benzyloxycarbonyl-Phe-Arg-aminomethyl coumarin
substrate inhibition
biotin-Phe-Ala-fluoromethylketone
-
chicken cystatin
-
-
guanidine hydrochloride
activates at low concentrations but inhibits above 2 M
heparan sulfate
-
Human cystatin C
-
-
KSCN
activates at low concentrations but inhibits above 1 M
L-kininogen
-
L-trans-epoxysuccinylleucylamido(4-guanidino)butane
strong inhibitor
N-carbobenzoxy-FA-diazomethylketone
inhibition in vivo
N-carbobenzoxy-FF-diazomethylketone
inhibition in vivo
N-carbobenzoxy-FK-acyloxymethylketone
inhibition in vivo
N-carbobenzoxy-VVR-4-nitroanilide
strong substrate inhibition
pregnancy zone protein
50% inhibition at a 1:1 ratio of cruzipain and pregnancy zone protein
-
Urea
70% inhibition at 5 M
[(S)-1-carboxy-2-phenylethyl]-carbamoyl-L-Arg-L-Val-arginal
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
guanidine hydrochloride
activates at low concentrations but inhibits above 2 M
KSCN
activates at low concentrations but inhibits above 1 M
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0003 - 0.001
2-aminobenzoic acid-KLRFSKQ-N-(2,4-dinitrophenyl)ethylenediamine
0.000065 - 0.00012
2-aminobenzoyl-LGMISLMKRPQ-N-(2,4-dinitrophenyl)ethylenediamine
0.028 - 0.029
azocasein
-
0.043
benzoyl-His-Arg-7-amido-4-methylcoumarin
pH 6.3, 37°C
-
0.066
benzoyl-PFR-4-nitroanilide
0.0006 - 0.015
benzoyl-Phe-Arg-7-amido-4-methylcoumarin
-
0.0119
benzoyl-RR-aminomethyl coumarin
pH 6.3, 37°C
0.001
benzoyl-Tyr-Arg-7-amido-4-methylcoumarin
-
-
0.0011
benzyloxycarbonyl-FR-aminomethyl coumarin
pH 6.3, 37°C
0.0008 - 0.0032
benzyloxycarbonyl-Phe-Arg-aminomethyl coumarin
0.011
N-carbobenzoxy-Arg-Arg-2-(4-methoxy)-naphthylamide
-
0.033
N-carbobenzoxy-VVR-4-nitroanilide
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0683 - 0.08
2-aminobenzoic acid-KLRFSKQ-N-(2,4-dinitrophenyl)ethylenediamine
0.025 - 0.0583
2-aminobenzoyl-LGMISLMKRPQ-N-(2,4-dinitrophenyl)ethylenediamine
0.172
benzoyl-His-Arg-7-amido-4-methylcoumarin
-
-
0.0433 - 0.193
benzoyl-Phe-Arg-7-amido-4-methylcoumarin
-
0.0783
benzoyl-RR-aminomethyl coumarin
-
0.158
benzyloxycarbonyl-FR-aminomethyl coumarin
-
2.16 - 4.49
benzyloxycarbonyl-Phe-Arg-aminomethyl coumarin
0.0805
N-carbobenzoxy-Arg-Arg-2-(4-methoxy)-naphthylamide
-
additional information
additional information
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.48 - 0.79
azocasein
-
0.023 - 0.1
benzyloxycarbonyl-Phe-Arg-aminomethyl coumarin
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.008 - 1.68
4-nitroanilide derivatives
0.1 - 3.77
beta-naphthylamide derivatives
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
assay with azocasein
7
beta-naphthylamide derivatives as substrate
7.6
assay with 4-nitroanilide
9
4-nitroanilide derivatives as substrate
additional information
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 7.4
hydrophobic substrates reach 50% activity at pH 4.5 and maintain full activity above pH 7.5, more hydrophilic side chains only reach 50% activity at pH 5.5-6
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
Q598P5_TRYCR
467
0
49836
TrEMBL
-
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
23540
electrospray mass spectrometry
35000 - 60000
SDS-PAGE, depending on electrophoretic conditions
36300
calculated from nucleotide sequence
43000
deglycosylated enzyme, SDS-PAGE
51000
glycosylated enzyme, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
bound to benzoyl-Arg-Ala-fluoromethyl ketone and benzoyl-Tyr-Ala-fluoromethyl ketone, hanging drop vapor diffusion method
complexed with benzyloxycarbonyl-Phe-Ala-fluoromethylketone
enzyme lacking the C-terminal domain, complexed with benzyloxycarbonyl-Phe-Ala-fluoromethylketone
recombinant protein and truncated version
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C25A
enzyme is inactive
E219P
variant with altered cleavage recognition site
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2.75
exponential decrease of activity in the presence of substrate
638891
8.7
unstable in the absence of substrate at 37°C
638878
additional information
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
unstable in the absence of substrate at pH 8.7
45
E219P mutant, stable for 10 h
5 - 37
inactivation at pH 2.25 and pH 10 increases with increasing temperature
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
unstable in the absence of substrate at pH 8.7 and 37°C
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
active complete mature enzyme
enzyme and truncated version
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Alvarez, V.; Parussini, F.; Aslund, L.; Cazzulo, J.J.
Expression in insect cells of active mature cruzipain from Trypanosoma cruzi, containing its C-terminal domain
Protein Expr. Purif.
26
467-475
2002
Trypanosoma cruzi (Q598P5), Trypanosoma cruzi
Manually annotated by BRENDA team
Alves, L.C.; Melo, R.L.; Cezari, M.H.S.; Sanderson, S.J.; Mottram, J.C.; Coombs, G.H.; Juliano, L.; Juliano, M.A.
Analysis of the S2 subsite specificities of the recombinant cysteine proteinases CPB of Leishmania mexicana, and cruzain of Trypanosoma cruzi, using fluorescent substrates containing non-natural basic amino acids
Mol. Biochem. Parasitol.
117
137-143
2001
Trypanosoma cruzi (Q598P5), Trypanosoma cruzi
Manually annotated by BRENDA team
Cazzulo, J.J.; Bravo, M.; Raimond, A.; Engstroem, U.; Lindeberg, G.; Hellman, U.
Hydrolysis of synthetic peptides by cruzipain, the major cysteine proteinase from Trypanosoma cruzi, provides evidence for self-processing and the possibility of more specific substrates for the enzyme
Cell. Mol. Biol.
42
691-696
1996
Trypanosoma cruzi (Q598P5), Trypanosoma cruzi
Manually annotated by BRENDA team
Cazzulo, J.J.; Cazzulo Franke, M.C.; Martinez, J.; Franke de Cazzulo, B.M.
Some kinetic properties of a cysteine proteinase (cruzipain) from Trypanosoma cruzi
Biochim. Biophys. Acta
1037
186-191
1990
Trypanosoma cruzi (Q598P5)
Manually annotated by BRENDA team
Cazzulo, J.J.; Stoka, V.; Turk, V.
Cruzipain, the major cysteine proteinase from the protozoan parasite Trypanosoma cruzi
Biol. Chem.
378
1-10
1997
Trypanosoma cruzi (Q598P5), Trypanosoma cruzi
Manually annotated by BRENDA team
Del Nery, E.; Juliano, M.A.; Meldal, M.; Svendsen, I.; Scharfstein, J.; Walmsley, A.; Juliano, L.
Characterization of the substrate specificity of the major cysteine protease (cruzipain) from Trypanosoma cruzi using a portion-mixing combinatorial library and fluorogenic peptides
Biochem. J.
323
427-433
1997
Trypanosoma cruzi (Q598P5)
-
Manually annotated by BRENDA team
Eakin, A.E.; McGrath, M.E.; McKerrow, J.H.; Fletterick, R.J.; Craik, C.S.
Production of crystallizable cruzain, the major cysteine protease from Trypanosoma cruzi
J. Biol. Chem.
268
6115-6118
1993
Trypanosoma cruzi (Q598P5), Trypanosoma cruzi
Manually annotated by BRENDA team
Gillmor, S.A.; Craik, C.S.; Fletterick, R.J.
Structural determinants of specificity in the cysteine protease cruzain
Protein Sci.
6
1603-1611
1997
Trypanosoma cruzi (Q598P5), Trypanosoma cruzi
Manually annotated by BRENDA team
Judice, W.A.; Cezari, M.H.; Lima, A.P.; Scharfstein, J.; Chagas, J.R.; Tersariol, I.L.; Juliano, M.A.; Juliano, L.
Comparison of the specificity, stability and individual rate constants with respective activation parameters for the peptidase activity of cruzipain and its recombinant form, cruzain, from Trypanosoma cruzi
Eur. J. Biochem.
268
6578-6586
2001
Trypanosoma cruzi (Q598P5), Trypanosoma cruzi
Manually annotated by BRENDA team
Lima, A.P.C.A.; Almeida, P.C.; Tersariol, I.L.S.; Schmitz, V.; Schmaier, A.H.; Juliano, L.; Hirata, I.Y.; Muller-Esterl, W.; Chagas, J.R.; Scharfstein, J.
Heparan sulfate modulates kinin release by Trypanosoma cruzi through the activity of cruzipain
J. Biol. Chem.
277
5875-5881
2002
Trypanosoma cruzi (Q598P5), Trypanosoma cruzi
Manually annotated by BRENDA team
McGrath, M.E.; Eakin, A.E.; Engel, J.C.; McKerrow, J.H.; Craik, C.C.; Fletterick, R.J.
The crystal structure of cruzain: a therapeutic target for Chagas' disease
J. Mol. Biol.
247
251-259
1995
Trypanosoma cruzi (Q598P5), Trypanosoma cruzi
Manually annotated by BRENDA team
Ramos, A.M.; Duschak, V.G.; Gerez de Burgos, N.M.; Barboza, M.; Remedi, M.S.; Vides, M.A.; Chiabrando, G.A.
Trypanosoma cruzi: cruzipain and membrane-bound cysteine proteinase isoform(s) interacts with human alpha2-macroglobulin and pregnancy zone protein
Exp. Parasitol.
100
121-130
2002
Trypanosoma cruzi (Q598P5), Trypanosoma cruzi
Manually annotated by BRENDA team
Salvati, L.; Mattu, M.; Polticelli, F.; Tiberi, F.; Gradoni, L.; Venturini, G.; Bolognesi, M.; Ascenzi, P.
Modulation of the catalytic activity of cruzipain, the major cysteine proteinase from Trypanosoma cruzi, by temperature and pH
Eur. J. Biochem.
268
3253-3258
2001
Trypanosoma cruzi (Q598P5), Trypanosoma cruzi
Manually annotated by BRENDA team
Schnapp, A.R.; Eickhoff, C.S.; Sizemore, D.; Curtiss, R.3rd.; Hoft, D.F.
Cruzipain induces both mucosal and systemic protection against Trypanosoma cruzi in mice
Infect. Immun.
70
5065-5074
2002
Trypanosoma cruzi (Q598P5), Trypanosoma cruzi
Manually annotated by BRENDA team
Sterin-Borda, L.; Giordanengo, L.; Joensen, L.; Gea, S.
Cruzipain induces autoantibodies against cardiac muscarinic acetylcholine receptors. Functional and pathological implications
Eur. J. Immunol.
33
2459-2468
2003
Trypanosoma cruzi (Q598P5), Trypanosoma cruzi
Manually annotated by BRENDA team
Stoka, V.; Turk, B.; McKerrow, J.H.; Bjork, I.; Cazzulo, J.J.; Turk, V.
The high stability of cruzipain against pH-induced inactivation is not dependent on its C-terminal domain
FEBS Lett.
469
29-32
2000
Trypanosoma cruzi (Q598P5)
Manually annotated by BRENDA team
Cazzulo, J.J.; Martinez, J.; Parodi, A.J.; Wernstedt, C.; Hellman, U.
On the post-translational modifications at the C-terminal domain of the major cysteine proteinase (cruzipain) from Trypanosoma cruzi
FEMS Microbiol. Lett.
79
411-416
1992
Trypanosoma cruzi (Q598P5), Trypanosoma cruzi
Manually annotated by BRENDA team