Molecular characterization and determination of the biochemical properties of cathepsin L of Trichinella spiralis

Liu, Ruo Dan; Meng, Xiang Yu; Li, Chen Le; Long, Stuart A.; Cui, Jing; Wang, Zhong Quan

doi:10.1186/s13567-022-01065-6

Cited by 11 publications

(17 citation statements)

References 78 publications

(86 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Phylogenetic analysis indicated that Dd-CPL-1 has the closest evolutionary phylogenetic relationship with the ortholog from the free-living fungivorous nematode A. avenae sharing 75.34% amino acid sequence identity. Molecular modelling showed that the structure of Dd-CPL-1 is composed of two globular domains divided by an active-site cleft, which is quite similar to that of other nematodes, such as B. xylophilus 36 , Angiostrongylus cantonensis 37 and Trichinella spiralis 38 . These results suggest that the CPL genes are conserved across the nematode phylum and Dd-CPL-1 may share similar functions with its orthologs in other nematode species.…”

Section: Discussionmentioning

confidence: 86%

Silencing Ditylenchus destructor cathepsin L-like cysteine protease has negative pleiotropic effect on nematode ontogenesis

Huang,

Cong,

Liu

et al. 2024

Sci Rep

View full text Add to dashboard Cite

Ditylenchus destructor is a migratory plant-parasitic nematode that severely harms many agriculturally important crops. The control of this pest is difficult, thus efficient strategies for its management in agricultural production are urgently required. Cathepsin L-like cysteine protease (CPL) is one important protease that has been shown to participate in various physiological and pathological processes. Here we decided to characterize the CPL gene (Dd-cpl-1) from D. destructor. Analysis of Dd-cpl-1 gene showed that Dd-cpl-1 gene contains a signal peptide, an I29 inhibitor domain with ERFNIN and GNFD motifs, and a peptidase C1 domain with four conserved active residues, showing evolutionary conservation with other nematode CPLs. RT-qPCR revealed that Dd-cpl-1 gene displayed high expression in third-stage juveniles (J3s) and female adults. In situ hybridization analysis demonstrated that Dd-cpl-1 was expressed in the digestive system and reproductive organs. Silencing Dd-cpl-1 in 1-cell stage eggs of D. destructor by RNAi resulted in a severely delay in development or even in abortive morphogenesis during embryogenesis. The RNAi-mediated silencing of Dd-cpl-1 in J2s and J3s resulted in a developmental arrest phenotype in J3 stage. In addition, silencing Dd-cpl-1 gene expression in female adults led to a 57.43% decrease in egg production. Finally, Dd-cpl-1 RNAi-treated nematodes showed a significant reduction in host colonization and infection. Overall, our results indicate that Dd-CPL-1 plays multiple roles in D. destructor ontogenesis and could serve as a new potential target for controlling D. destructor.

show abstract

Section: Discussionmentioning

confidence: 86%

Silencing Ditylenchus destructor cathepsin L-like cysteine protease has negative pleiotropic effect on nematode ontogenesis

Huang,

Cong,

Liu

et al. 2024

Sci Rep

View full text Add to dashboard Cite

show abstract

“…In Giardia duodenalis, cysteine proteases play a role in the pathophysiology of parasitic infection by inducing the degradation and redistribution of the intestinal epithelial cytoskeletal protein villin via myosin light chain kinase (29). Also in Trichinella spiralis, cysteine proteases are important for the nutrient acquisition, immune evasion, and invasion in the host (30). In Entamoeba histolytica with a closer in origin to H. meleagridis, cysteine proteases are important virulence factors that help them to disrupt the host intestinal epithelial barrier functions and circumvent host immune responses (31)(32)(33).…”

Section: Discussionmentioning

confidence: 99%

Tandem mass tag-based quantitative proteomics analyses of a chicken-original virulent and its attenuated Histomonas meleagridis strain in China

et al. 2023

View full text Add to dashboard Cite

IntroductionHistomonas meleagridis can cause histomonosis in poultry. Due to the prohibition of effective drugs, the prevention and treatment of the disease requires new strategies. Questions about its pathogenic mechanisms and virulence factors remain puzzling.MethodsTo address these issues, a tandem mass tag (TMT) comparative proteomic analysis of a virulent strain and its attenuated strain of Chinese chicken-origin was performed.ResultsA total of 3,494 proteins were identified in the experiment, of which 745 proteins were differentially expressed (fold change ≥1.2 or ≤0.83 and p < 0.05), with 192 up-regulated proteins and 553 down-regulated proteins in the virulent strain relative to the attenuated strain.DiscussionSurface protein BspA like, digestive cysteine proteinase, actin, and GH family 25 lysozyme were noted among the proteins up regulated in virulent strains, and these several proteins may be directly related to the pathogenic capacity of the histomonad. Ferredoxin, 60S ribosomal protein L6, 40S ribosomal protein S3, and NADP-dependent malic enzyme which associated with biosynthesis and metabolism were also noted, which have the potential to be new drug targets. The up-regulation of alpha-amylase, ras-like protein 1, ras-like protein 2, and involucrin in attenuated strains helps to understand how it is adapted to the long-term in vitro culture environment. The above results provide some candidate protein-coding genes for further functional verification, which will help to understand the molecular mechanism of pathogenicity and attenuation of H. meleagridis more comprehensively.

show abstract

“…The pMAL-c2X/TsCatL2 recombinant plasmid were stored in our laboratory. Recombinant proteins (rTsCatL2) and MBP were expressed in Escherichia coli Rosetta-gami B (DE3), purified with amylose resin, and detected in previous studies [ 14 ]. To eliminate endotoxin interference in rTsCatL2, endotoxin removal resin (Thermo Fisher, USA) was used.…”

Section: Methodsmentioning

confidence: 99%

“…To eliminate endotoxin interference in rTsCatL2, endotoxin removal resin (Thermo Fisher, USA) was used. Mouse anti-rTsACatL2 serum, anti-rTsASP2 serum and anti-MBP serum were produced by immunizing BALB/c mice with rTsCatL/rTsASP2/MBP mixed with adjuvant via intramuscular injection and collecting the blood of the mice to isolate the serum [ 6 , 14 ]. The human colonic epithelial cell line Caco-2 and the mouse myogenic cell line C2C12 were obtained from the Cell Resource Centre, Chinese Academy of Sciences.…”

Section: Methodsmentioning

confidence: 99%

Trichinella spiralis cathepsin L damages the tight junctions of intestinal epithelial cells and mediates larval invasion

Liu,

Meng,

et al. 2023

PLoS Negl Trop Dis

Self Cite

View full text Add to dashboard Cite

Background Cathepsin L, a lysosomal enzyme, participates in diverse physiological processes. Recombinant Trichinella spiralis cathepsin L domains (rTsCatL2) exhibited natural cysteine protease activity and hydrolyzed host immunoglobulin and extracellular matrix proteins in vitro, but its functions in larval invasion are unknown. The aim of this study was to explore its functions in T. spiralis invasion of the host’s intestinal epithelial cells. Methodology/principal findings RNAi significantly suppressed the expression of TsCatL mRNA and protein with TsCatL specific siRNA-302. T. spiralis larval invasion of Caco-2 cells was reduced by 39.87% and 38.36%, respectively, when anti-TsCatL2 serum and siRNA-302 were used. Mice challenged with siRNA-302-treated muscle larvae (ML) exhibited a substantial reduction in intestinal infective larvae, adult worm, and ML burden compared to the PBS group, with reductions of 44.37%, 47.57%, and 57.06%, respectively. The development and fecundity of the females from the mice infected with siRNA-302-treated ML was significantly inhibited. After incubation of rTsCatL2 with Caco-2 cells, immunofluorescence test showed that the rTsCatL2 gradually entered into the cells, altered the localization of cellular tight junction proteins (claudin 1, occludin and zo-1), adhesion junction protein (e-cadherin) and extracellular matrix protein (laminin), and intercellular junctions were lost. Western blot showed a 58.65% reduction in claudin 1 expression in Caco-2 cells treated with rTsCatL2. Co-IP showed that rTsCatL2 interacted with laminin and collagen I but not with claudin 1, e-cadherin, occludin and fibronectin in Caco-2 cells. Moreover, rTsCatL2 disrupted the intestinal epithelial barrier by inducing cellular autophagy. Conclusions rTsCatL2 disrupts the intestinal epithelial barrier and facilitates T. spiralis larval invasion.

show abstract

Molecular characterization and determination of the biochemical properties of cathepsin L of Trichinella spiralis

Cited by 11 publications

References 78 publications

Silencing Ditylenchus destructor cathepsin L-like cysteine protease has negative pleiotropic effect on nematode ontogenesis

Silencing Ditylenchus destructor cathepsin L-like cysteine protease has negative pleiotropic effect on nematode ontogenesis

Tandem mass tag-based quantitative proteomics analyses of a chicken-original virulent and its attenuated Histomonas meleagridis strain in China

Trichinella spiralis cathepsin L damages the tight junctions of intestinal epithelial cells and mediates larval invasion

Contact Info

Product

Resources

About