Molecular cloning and characterization of a cathepsin L-like cysteine protease of Angiostrongylus cantonensis

Bai, Huifang; Cao, Yizhen; Chen, Yunqiu; Zhang, Lingmin; Wu, Cheng-Shong; Zhan, Xi; Cheng, Minquan

doi:10.1016/j.ijbiomac.2019.10.243

Cited by 4 publications

(2 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The inhibitor_I29 domain of TsCatL is a prepeptide that is removed by self-hydrolysis under acidic conditions, and the Pept_C1 domain becomes the mature peptide with enzymatic activity. This result is consistent with previous findings that cathepsin L proenzymes can autocatalytically cleave under acidic conditions in vitro [61][62][63]. Cysteine proteases usually exist as preproenzymes that can be self-hydrolysed under acidic conditions, removing the precursor peptide and transforming into an active mature enzyme [64].…”

Section: Discussionsupporting

confidence: 92%

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

Liu

Meng

et al. 2022

Vet Res

View full text Add to dashboard Cite

Cathepsin L is an important cysteine protease, but its function in T. spiralis remains unclear. The aim of this research was to explore the biological characteristics of T. spiralis cathepsin L (TsCatL) and its role in T. spiralis-host interactions. Bioinformatic analysis revealed the presence of the cysteine protease active site residues Gln, Cys, His and Asn in mature TsCatL, as well as specific motifs of cathepsin L similar to ERFNIN and GYLND in the prepeptide of TsCatL. Molecular docking of mature TsCatL and E64 revealed hydrophobic effects and hydrogen bonding interactions. Two domains of TsCatL (TsCatL2) were cloned and expressed, and recombinant TsCatL2 (rTsCatL2) was autocatalytically cleaved under acidic conditions to form mature TsCatL. TsCatL was transcribed and expressed in larvae and adults and located in the stichosome, gut and embryo. Enzyme kinetic tests showed that rTsCatL2 degraded the substrate Z-Phe-Arg-AMC under acidic conditions, which was inhibited by E64 and PMSF and enhanced by EDTA, L-cysteine and DTT. The kinetic parameters of rTsCatL2 were a Km value of 48.82 μM and Vmax of 374.4 nM/min at pH 4.5, 37 °C and 5 mM DTT. In addition, it was shown that rTsCatL2 degraded haemoglobin, serum albumin, immunoglobulins (mouse IgG, human IgG and IgM) and extracellular matrix components (fibronectin, collagen I and laminin). The proteolytic activity of rTsCatL2 was host specific and significantly inhibited by E64. rTsCatL2 possesses the natural activity of a sulfhydryl-containing cysteine protease, and TsCatL is an important digestive enzyme that seems to be important for the nutrient acquisition, immune evasion and invasion of Trichinella in the host.

show abstract

Section: Discussionsupporting

confidence: 92%

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

Liu

Meng

et al. 2022

Vet Res

View full text Add to dashboard Cite

show abstract

“…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: 85%

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

Molecular characterization of a novel cathepsin L from Trichinella spiralis and its participation in invasion, development and reproduction

et al. 2021

View full text Add to dashboard Cite

Molecular cloning and characterization of a cathepsin L-like cysteine protease of Angiostrongylus cantonensis

Cited by 4 publications

References 35 publications

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

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

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

Molecular characterization of a novel cathepsin L from Trichinella spiralis and its participation in invasion, development and reproduction

Contact Info

Product

Resources

About