Processing of the Papain Precursor

Vernet, Thierry; Berti, Paul J.; Montigny, Chantal de; Musil, Roy; Tessier, Daniel C.; Ménard, Robert; Magny, Marie-Claude; Storer, Andrew C.; Thomas, David Y.

doi:10.1074/jbc.270.18.10838

Cited by 135 publications

(60 citation statements)

References 47 publications

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“…We have shown that acidification of ricinosomes in vitro results in removal of the N-terminal propeptide within a few minutes, leading to the mature protease with the correct amino terminus and cleavage of the C terminus, removing the KDEL motif. We suggest that this maturation is due to autocleavage, as is the case for papain and other cysteine proteases (22), but cannot exclude the possibility of activation by low levels of another endoprotease. Loss of ricinosome integrity in vivo may occur with the acidification of the cytoplasm resulting from a change in tonoplast permeability at a late stage of PCD.…”

Section: Discussionmentioning

confidence: 99%

The ricinosomes of senescing plant tissue bud from the endoplasmic reticulum

Schmid

Simpson

Sarioglu

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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The ricinosome (synonym, precursor protease vesicle) is a novel organelle, found so far exclusively in plant cells. Electron microscopic studies suggest that it buds off from the endoplasmic reticulum in senescing tissues. Biochemical support for this unusual origin now comes from the composition of the purified organelle, which contains large amounts of a 45-kDa cysteine endoprotease precursor with a C-terminal KDEL motif and the endoplasmic reticulum lumen residents BiP (binding protein) and protein disulfide isomerase. Western blot analysis, peptide sequencing, and mass spectrometry demonstrate retention of KDEL in the protease proform. Acidification of isolated ricinosomes causes castor bean cysteine endopeptidase activation, with cleavage of the N-terminal propeptide and the C-terminal KDEL motif. We propose that ricinosomes accumulate during senescence by programmed cell death and are activated by release of protons from acidic vacuoles.Ricinus communis ͉ papain-type KDEL peptidase P rogrammed cell death (PCD) in higher plants and animals fulfills multiple functions in the differentiation or turnover of tissues and is triggered by pathogens at and around the site of infection as a mechanism to prevent proliferation of the disease (1, 2). Examples for elimination by PCD of organs that serve temporary functions are tadpole tails during metamorphosis and the endosperm of the castor bean during germination. In plants, the PCD of whole organs such as leaves, flowers, and fruits is designated as senescence (3).Biochemical pathways accompanying PCD or apoptosis in mammals have been elucidated in detail (reviewed in ref. 4). Plant cells undergoing senescence by PCD express papain-type cysteine endoproteases with a C-terminal KDEL sequence (5). They attract attention because they are present in the senescing endosperm cells of castor bean (Ricinus communis) as relatively inactive 45-kDa proenzymes until the final stages of PCD, at which time they are processed into the 35-kDa mature form with a 50-100 times higher activity (6).The cysteine endopeptidase from senescing castor bean endosperm (CysEP) is homologous to papain-type KDEL cysteine endopeptidases from senescing flower petals of daylily, the maturing French bean pods, and the cotyledons of germinating mung bean and vetch (6). In castor bean, daylily, and mung bean, the KDEL-tailed propeptidase is stored in a special organelle, called ricinosome (5, 6) or KDEL-tailed cysteine proteinaseaccumulating vesicles (KV) (7). Ricinosomes were first described and ultrastructurally characterized in the endosperm of R. communis (8, 9), whereas KV were described as cytoplasmic ''foci'' in Vigna radiata (10). The mature CysEP was originally purified from germinating castor bean endosperm (11). Sequencing of cDNA clones from the endosperm of germinating seedlings and from developing seeds established the presence of a presequence for cotranslational targeting into the lumen of the endoplasmic reticulum (ER), an N-terminal propeptide and a C-terminal KDEL motif. Immunogold l...

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Section: Discussionmentioning

confidence: 99%

The ricinosomes of senescing plant tissue bud from the endoplasmic reticulum

Schmid

Simpson

Sarioglu

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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“…Intermolecular processing was studied by generating an inactive procathepsin L mutant, where the catalytic Cys residue at position 26 was substituted with a Gly, which was incapable of autoactivation. Our results show that P. pastoris-expressed cathepsin L can autoactivate at low pH to an active intermediate form by an initial cleavage within the conserved heptapeptide (Gly-Xaa-Asn-Xaa-PheXaa-Asp), as previously described for papain (32). Further activation of the enzyme occurs by cleavage in the vicinity of the N terminus.…”

mentioning

confidence: 63%

“…It was suggested that the lowering of the pH perturbed the negative charge of Asp Ϫ36 resulting in a conformational change that switched on the processing events by allowing proteolysis to occur at the Ala Ϫ37 /Asp Ϫ36 bond. Following this primary cleavage further removal of the remaining amino acids of the propeptide may result from the proteolytic activity of the intermediate species, resulting in fully active mature protease (32).…”

mentioning

confidence: 99%

“…Earlier studies on the processing of yeast-expressed recombinant papain identified a conserved heptapeptide (Gly-XaaAsn-Xaa-Phe-Xaa-Asp) motif located between residues Ϫ42 and Ϫ36 in the propeptide that may be a site of initial cleavage in the pH-dependent autoactivation of the enzyme (32). It was suggested that the lowering of the pH perturbed the negative charge of Asp Ϫ36 resulting in a conformational change that switched on the processing events by allowing proteolysis to occur at the Ala Ϫ37 /Asp Ϫ36 bond.…”

mentioning

confidence: 99%

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Cathepsin L1, the Major Protease Involved in Liver Fluke (Fasciola hepatica) Virulence

Collins

Stack

O’Neill

et al. 2004

Journal of Biological Chemistry

146

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The secretion and activation of the major cathepsin L1 cysteine protease involved in the virulence of the helminth pathogen Fasciola hepatica was investigated. Only the fully processed and active mature enzyme can be detected in medium in which adult F. hepatica are cultured. However, immunocytochemical studies revealed that the inactive procathepsin L1 is packaged in secretory vesicles of epithelial cells that line the parasite gut. These observations suggest that processing and activation of procathepsin L1 occurs following secretion from these cells into the acidic gut lumen. Expression of the 37-kDa procathepsin L1 in Pichia pastoris showed that an intermolecular processing event within a con-

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“…The sequence motif GXNXFXD is highly conserved in Sv-CPL-1, although the Phe is replaced by Ile, as with other nematode CPLs. It occurs in proregions of cysteine proteases of the papain superfamily, and may serve intramolecular processing of papain (Vernet et al, 1995).…”

Section: Isolation and Characterisation Of Cathepsin L-like Cdnamentioning

confidence: 99%

A cathepsin L-like protease from Strongylus vulgaris: An orthologue of Caenorhabditis elegans CPL-1

Ultaigh

Carolan

Britton

et al. 2009

Experimental Parasitology

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a b s t r a c tCathespin L-like proteases (CPLs), characterized from a wide range of helminths, are significant in helminth biology. For example, in Caenorhabditis elegans CPL is essential for embryogenesis. Here, we report a cathepsin L-like gene from three species of strongyles that parasitize the horse, and describe the isolation of a cpl gene (Sv-cpl-1) from Strongylus vulgaris, the first such from equine strongyles. It encodes a protein of 354 amino acids with high similarity to other parasitic Strongylida (90-91%), and C. elegans CPL-1 (87%), a member of the same Clade. As S. vulgaris cpl-1 rescued the embryonic lethal phenotype of the C. elegans cpl-1 mutant, these genes may be orthologues, sharing the same function in each species. Targeting Sv-CPL-1 might enable novel control strategies by decreasing parasite development and transmission.

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Processing of the Papain Precursor

Cited by 135 publications

References 47 publications

The ricinosomes of senescing plant tissue bud from the endoplasmic reticulum

The ricinosomes of senescing plant tissue bud from the endoplasmic reticulum

Cathepsin L1, the Major Protease Involved in Liver Fluke (Fasciola hepatica) Virulence

A cathepsin L-like protease from Strongylus vulgaris: An orthologue of Caenorhabditis elegans CPL-1

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