Molecular cloning and characterization of cathepsin L encoding genes from Fasciola gigantica

Grams, Rudi; Vichasri-Grams, Suksiri; Sobhon, Prasert; Upatham, E. S.; Viyanant, Vithoon

doi:10.1016/s1383-5769(01)00068-x

Cited by 60 publications

(25 citation statements)

References 25 publications

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“…However, in situ hybridisation studies by Grams et al (2001) with F. gigantica and by us with F. hepatica (Fig. 3) are consistent with immunolocalisation studies and show that the vast majority of cathepsin L proteases are expressed in the cells lining the parasite's guts.…”

Section: The Cathepsin L Gene Familysupporting

confidence: 82%

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Fasciola hepatica cathepsin L-like proteases: biology, function, and potential in the development of first generation liver fluke vaccines

Dalton

Neill

Stack

et al. 2003

International Journal for Parasitology

235

165

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Fasciola hepatica secretes cathepsin L proteases that facilitate the penetration of the parasite through the tissues of its host, and also participate in functions such as feeding and immune evasion. The major proteases, cathepsin L1 (FheCL1) and cathepsin L2 (FheCL2) are members of a lineage that gave rise to the human cathepsin Ls, Ks and Ss, but while they exhibit similarities in their substrate specificities to these enzymes they differ in having a wider pH range for activity and an enhanced stability at neutral pH. There are presently 13 Fasciola cathepsin L cDNAs deposited in the public databases representing a gene family of at least seven distinct members, although the temporal and spatial expression of each of these members in the developmental stage of F. hepatica remains unclear. Immunolocalisation and in situ hybridisation studies, using antibody and DNA probes, respectively, show that the vast majority of cathepsin L gene expression is carried out in the epithelial cells lining the parasite gut. Within these cells the enzyme is packaged into secretory vesicles that release their contents into the gut lumen for the purpose of degrading ingested host tissue and blood. Liver flukes also express a novel multi-domain cystatin that may be involved in the regulation of cathepsin L activity. Vaccine trials in both sheep and cattle with purified native FheCL1 and FheCL2 have shown that these enzymes can induce protection, ranging from 33 to 79%, to experimental challenge with metacercariae of F. hepatica, and very potent anti-embryonation/hatch rate effects that would block parasite transmission. In this article we review the vaccine trials carried out over the past 8 years, the role of antibody and T cell responses in mediating protection and discuss the prospects of the cathepsin Ls in the development of first generation recombinant liver fluke vaccines. q

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Section: The Cathepsin L Gene Familysupporting

confidence: 82%

“…The number of F. gigantica cathepsin L entries in the databases has also grown mainly due to the efforts of Grams et al (2001) and Yamasaki et al (2002), and unsurprisingly many of these genes appear to be homologues of the F. hepatica genes. The family as it stands is presented in Fig.…”

Section: The Cathepsin L Gene Familymentioning

confidence: 99%

Fasciola hepatica cathepsin L-like proteases: biology, function, and potential in the development of first generation liver fluke vaccines

Dalton

Neill

Stack

et al. 2003

International Journal for Parasitology

235

165

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“…Fourteen of the Fasciola cathepsin L sequences were not fulllength cDNAs and/or differed by only a small number of nucleotides and thus likely represent different alleles rather than individual genes (12,21). This is possible, and perhaps likely, given the occurrence of triploid fasciolids (with an extra allele available) in both temperate and tropical regions (22)(23)(24).…”

Section: Phylogenetic Analysis Of Fasciola Cathepsin L Sequences-mentioning

confidence: 99%

“…This is possible, and perhaps likely, given the occurrence of triploid fasciolids (with an extra allele available) in both temperate and tropical regions (22)(23)(24). However, Grams et al (21) estimated from Southern blot analysis that at least 10 cathepsin L genes, formed by duplication events, exist in F. gigantica, but that other, more divergent sequences would not have been detected as a result of the stringent hybridization conditions used. Until significant genomic sequence information is available for a member of the genus, the contribution of allelic variation to the current repertoire of Fasciola cathepsin Ls remains to be fully determined.…”

Section: Phylogenetic Analysis Of Fasciola Cathepsin L Sequences-mentioning

confidence: 99%

Proteomics and Phylogenetic Analysis of the Cathepsin L Protease Family of the Helminth Pathogen Fasciola hepatica

Robinson

Tort

Lowther

et al. 2008

Molecular & Cellular Proteomics

120

131

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Cathepsin L proteases secreted by the helminth pathogen Fasciola hepatica have functions in parasite virulence including tissue invasion and suppression of host immune responses. Using proteomics methods alongside phylogenetic studies we characterized the profile of cathepsin L proteases secreted by adult F. hepatica and hence identified those involved in host-pathogen interaction. Phylogenetic analyses showed that the Fasciola cathepsin L gene family expanded by a series of gene duplications followed by divergence that gave rise to three clades associated with mature adult worms (Clades 1, 2, and 5) and two clades specific to infective juvenile stages (Clades 3 and 4). Consistent with these observations our proteomics studies identified representatives from Clades 1, 2, and 5 but not from Clades 3 and 4 in adult F. hepatica secretory products. Clades 1 and 2 account for 67.39 and 27.63% of total secreted cathepsin Ls, respectively, suggesting that their expansion was positively driven and that these proteases are most critical for parasite survival and adaptation. Sequence comparison studies revealed that the expansion of cathepsin Ls by gene duplication was followed by residue changes in the S2 pocket of the active site. Our biochemical studies showed that these changes result in alterations in substrate binding and suggested that the divergence of the cathepsin L family produced a repertoire of enzymes with overlapping and complementary substrate specificities that could cleave host macromolecules more efficiently. Although the cathepsin Ls are produced as zymogens containing a prosegment and mature domain, all secreted enzymes identified by MS were processed to mature active enzymes. The prosegment region was highly conserved between the clades except at the boundary of prosegment and mature enzyme. Despite the lack of conservation at this section, sites for exogenous cleavage by asparaginyl endopeptidases and a Leu-Ser2His motif for autocatalytic cleavage by cathepsin Ls were preserved.

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“…On the other hand, drawbacks in the use of artificial carriers may originate from steric hindrances due to the close proximity of the antigenic peptides or to the type of the linkage between the carrier and epitope. 16,17 We herein report on the use of a peptide based on the H-Asp 110 -Lys-Ile-Asp-Trp-Arg-Glu-Ser-Gly-Tyr-Val-Thr-GluVal 123 -OH (Fas14p) sequence of F. gigantica cathepsin L-cysteine proteinase 18,19 in fasciolosis immunodiagnosis and prevention. According to Cornelissen et al 5 peptides of sequences 103-122 and 110-129 were verified to be the major immunodominant epitopes of cathepsin-L 1 .…”

Section: Introductionmentioning

confidence: 99%

Fasciola gigantica cathepsin L proteinase‐based synthetic peptide for immunodiagnosis and prevention of sheep fasciolosis

et al. 2008

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Molecular cloning and characterization of cathepsin L encoding genes from Fasciola gigantica

Cited by 60 publications

References 25 publications

Fasciola hepatica cathepsin L-like proteases: biology, function, and potential in the development of first generation liver fluke vaccines

Fasciola hepatica cathepsin L-like proteases: biology, function, and potential in the development of first generation liver fluke vaccines

Proteomics and Phylogenetic Analysis of the Cathepsin L Protease Family of the Helminth Pathogen Fasciola hepatica

Fasciola gigantica cathepsin L proteinase‐based synthetic peptide for immunodiagnosis and prevention of sheep fasciolosis

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