Molecular characterization and determination of the biochemical properties of cathepsin L of Trichinella spiralis.
Trichinella spiralis
cathepsin L
cysteine protease
enzymatic characterization
inhibitor
Journal
Veterinary research
ISSN: 1297-9716
Titre abrégé: Vet Res
Pays: England
ID NLM: 9309551
Informations de publication
Date de publication:
23 Jun 2022
23 Jun 2022
Historique:
received:
07
02
2022
accepted:
03
05
2022
entrez:
23
6
2022
pubmed:
24
6
2022
medline:
28
6
2022
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 35739604
doi: 10.1186/s13567-022-01065-6
pii: 10.1186/s13567-022-01065-6
pmc: PMC9229914
doi:
Substances chimiques
Immunoglobulin G
0
Cysteine Proteases
EC 3.4.-
Cathepsin L
EC 3.4.22.15
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
48Subventions
Organisme : National Natural Science Foundation of China
ID : 81802025
Organisme : National Natural Science Foundation of China
ID : 82172300
Organisme : Science and Technology Department of Henan Province
ID : 212102310146
Informations de copyright
© 2022. The Author(s).
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