Some kinetic properties of a cysteine proteinase (cruzipain) from Trypanosoma cruzi

Cazzulo, Juán José; Franke, M C Cazzulo; Martı́nez, Javier; Cazzulo, Berta M. Franke de

doi:10.1016/0167-4838(90)90166-d

Cited by 150 publications

(101 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The association rate constants for the interaction of cathepsin L and cruzipain with the Ii peptide are very similar, whereas the dissociation rate constant was considerably lower for cathepsin L ( Table 1). The fact that cruzipain is inhibited to about the same extent as cathepsin L, whereas related enzymes are inhibited only weekly or not at all [15], is in agreement with previous observations [11,13] that the active site geometry of cruzipain resembles mostly cathepsin L. The inhibitory li fragment shows no sequence homology with cystatins, but is homologous to thyroglobulin type-1 domain [19]. Recently, a protein from eggs of chum salmon has been isolated [20], which shares a high degree of homology with the thyroglobulin type-1 motif, and also inhibits cysteine proteinases.…”

Section: Discussionsupporting

confidence: 81%

See 1 more Smart Citation

A fragment of the major histocompatibility complex class II – associated p41 invariant chain inhibits cruzipain, the major cysteine proteinase from Trypanosoma cruzi

et al. 1997

Self Cite

View full text Add to dashboard Cite

A peptide fragment derived from the p41 form of the invariant chain (Ii) associated with the major histocompatibility complex (MHC) class II molecule has been shown to inhibit the mammalian lysosomal cysteine proteinase, cathepsin L, and to be a novel cysteine proteinase inhibitor, distinct from cystatins. Here we report that this same fragment also binds to and inhibits cruzipain, the cathepsin L-like enzyme from the protozoan parasite Trypanosoma cruzi. The binding of the Ii fragment to cruzipain is fast (k ass = 2.4 X10 7 M" 1 s" 1 ) and tight (Ki = 5.8 X10 n M). The inhibition is competitive. These results suggest the possibility of using the invariant chain as a model for the specific inhibition of cruzipain in vivo, i.e. as a potential drug to combat Chagas' disease.

show abstract

Section: Discussionsupporting

confidence: 81%

“…Cruzipain is inhibited by a range of organomercurial, epoxysuccinyl and peptide aldehyde compounds [10,11], as well as by protein inhibitors belonging to the cystatin superfamily [12,13]. The latter bind tightly to the enzyme with K\ values between 1 and 72 pM [12].…”

Section: Introductionmentioning

confidence: 99%

A fragment of the major histocompatibility complex class II – associated p41 invariant chain inhibits cruzipain, the major cysteine proteinase from Trypanosoma cruzi

et al. 1997

Self Cite

View full text Add to dashboard Cite

show abstract

“…Arg or Lys) in the PI position. This is a feature common to many mammalian and protozoan cysteine proteases (Pupkis and Coombs, 1984;Luaces and Barrett, 1988;Pamer et al, 1989;Cazzulo et al, 1990;Mason et al, 1985).…”

Section: Discussionmentioning

confidence: 99%

Characterisation of a cysteine protease from bloodstream forms of Trypanosoma congolense

Mbawa¹,

Gumm²,

Shaw

et al. 1992

European Journal of Biochemistry

View full text Add to dashboard Cite

A cysteine protease (trypanopain-Tc) with cathepsin-L-like properties has been purified from Trypanosoma congolense. The enzyme has an apparent molecular mass of 31 -3 2 kDa by SDS/ PAGE and 66 kDa by gel chromatography. It has a PI 7.4 and a high affinity for concanavalin A, Trypanopain-Tc catalyses the limited proteolysis of a variety of protein substrates such as fibrinogen, serum albumin and trypanosome variant-surface glycoprotein. It has minimal or no activity against casein or elastin.A variety of peptidyl amidomethylcoumarins and peptidyl diazomethanes were used to test the specificity of trypanopain-Tc. The better substrates had Arg or Lys in P1 and hydrophobic amino acids in P2 and P3. The best substrate found for trypanopain-Tc was Z-Phe-Arg-NHMec (Z, benzyloxycarbonyl ; NHMec, 7-amido-4-methylcoumarin). The kinetic constants for the hydrolysis, of ZPhe-Arg-NHMec were k,,, = 17.4 SKI, K, = 4.4 pM, kcal/Km = 4.0 pM-' ' s-', which are very similar to those of cathepsin L with this substrate. The specific substrates for cathepsin B (Z-ArgArg-NHMec) and cathepsin H (Arg-NHMec) were not hydrolysed by trypanopain-Tc under the conditions tested. The pH optimum of trypanopain-Tc against Z-Phe-Arg-NHMec was pH 6.0 but it showed a broad peak of activity extending well into the alkaline region. The enzyme was activated by low-molecular-mass thiol compounds and inhibited by cystatin, ~-trans-epoxysucciny1-4-guanidinobutane (E-64) and a variety of peptidyl diazomethanes. The most effective diazomethane inhibitors (Z-Leu-Leu-Met-CHN2, Z-Leu-Met-CHN2 and Z-Leu-Lys-CHN2, were inhibitory at nanomolar concentrations and were trypanocidal in vituo after 24 -48 h incubation in 2 20 pM [inhibitor]. However, it is not clear whether the trypanocidal activity of these inhibitors is a consequence of the inhibition of trypanopains or of some other essential proteolytic activities within the parasites.Cathepsins L and B play important roles in intracellular protein turnover (for review see Ballard, 1987;Marzella and Glaumann, 1987). They are also implicated in the degradation of extracellular matrix in diseases such as muscular dystrophy, emphysema and arthritis, and in association with tumour metastasis (Rochefort ct al., 1987;Kominami et al., 1987; Burnett and Stockley, 1985;Etherington et al., 1988). Recently, interest has been shown in the role of cysteine proteases in the pathology of parasitic infections. These proteases can be released from the parasites (Blum, 1976;Keene, 1986; Lockwood et al., 1988; Garber and Lemchuk-Favel, 1989;North et al., 1989; Boutignon et al., 1990) andmay beinvolved in the pathology of the diseases and the survival of the parasites within the host (Bremmer et al

show abstract

“…This glycoprotein of about 52-58 kDa has a C-terminal domain highly mannose glycosylated [6]. It has been reported that cruzipain is highly immunogenic in human infection [7,8].…”

Section: Introductionmentioning

confidence: 99%

Cruzipain, a major Trypanosoma cruzi antigen, conditions the host immune response in favor of parasite

Giordanengo¹,

Guiñazú²,

Stempin³

et al. 2002

Eur. J. Immunol.

View full text Add to dashboard Cite

We recently demonstrated that humoral immune response to cruzipain, a major antigen of Trypanosoma cruzi parasite, is implicated in the pathogenesis of experimental Chagas' disease. In the present study, the spleen cell phenotype and the cytokine profile induced by cruzipain in immunized mice were analyzed. The results showed that cruzipain increases the number of spleen cells with large size and granularity. Splenocyte populations with CD19 + , Mac-1 + , Gr-1 + and CD11c + positive surface markers significantly increased in immune mice compared to controls ones. Histological study revealed the presence of high number of megacariocyte and granulocyte-macrophage progenitors, indicating extramedullary hemopoiesis in spleens of immune mice. The finding of high levels of IL-4, IL5 and IL-10 and low levels of IFN-+ and IL-12 in supernatants of immune cells stimulated with cruzipain indicates a preferential activation of T2 type cells in immune animals. To investigate the role of innate immunity cells, the classical and alternative metabolic pathways of spleen macrophages from immune mice stimulated by cruzipain were also studied. The results showed an increase of urea associated with a decrease of nitrite levels, suggesting that cruzipain upregulates the arginase way. Therefore, cruzipain leads to T2 type cytokine profile which may enhance the arginase via in the macrophages promoting a susceptible mechanism to infection. Thus, we postulate that during T. cruzi infection, cruzipain could be used by the parasite to spread inside the host.

show abstract

Some kinetic properties of a cysteine proteinase (cruzipain) from Trypanosoma cruzi

Cited by 150 publications

References 11 publications

A fragment of the major histocompatibility complex class II – associated p41 invariant chain inhibits cruzipain, the major cysteine proteinase from Trypanosoma cruzi

A fragment of the major histocompatibility complex class II – associated p41 invariant chain inhibits cruzipain, the major cysteine proteinase from Trypanosoma cruzi

Characterisation of a cysteine protease from bloodstream forms of Trypanosoma congolense

Cruzipain, a major Trypanosoma cruzi antigen, conditions the host immune response in favor of parasite

Contact Info

Product

Resources

About