Molecular cloning and characterization of a cathepsin L-like cysteine protease of Angiostrongylus cantonensis
| Autor: | Yizhen Cao, Chunyun Wu, Lingmin Zhang, Mei Cheng, Huifang Bai, Yunqiu Chen, Ximei Zhan |
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| Rok vydání: | 2020 |
| Předmět: |
Models
Molecular Proteases Protein Conformation Cathepsin L medicine.medical_treatment 02 engineering and technology Biochemistry Microbiology Mice 03 medical and health sciences Structural Biology medicine Animals Amino Acid Sequence RNA Messenger Cloning Molecular Angiostrongylus Molecular Biology Gene Library 030304 developmental biology Cathepsin 0303 health sciences Protease Base Sequence biology Angiostrongylus cantonensis Gene Expression Regulation Developmental General Medicine 021001 nanoscience & nanotechnology biology.organism_classification medicine.disease Cysteine protease Protein Transport Angiostrongyliasis biology.protein Heterologous expression 0210 nano-technology |
| Zdroj: | International Journal of Biological Macromolecules. 153:1136-1146 |
| ISSN: | 0141-8130 |
| Popis: | Angiostrongylus cantonensis is a parasitic nematode dwelling in the heart and pulmonary arteries of rats, which can cause angiostrongyliasis in human by accidental infections, manifested as eosinophilic meningitis or meningoencephalitis. Cysteine proteases are the major class of endopeptidases that are expressed at a high level in A. cantonensis, which suggests it may play key roles in pathogenesis of the disease. In this study, the biological properties of the cathepsin L-like peptidase (Ac-cathL) of A. cantonensis were investigated. The Ac-cathL gene was identified from the fourth stage cDNA library of A. cantonensis, and then cloned and characterized by bioinformatics analysis and heterologous expression. The open reading frame (ORF) of Ac-cathL (1068 bp) encodes a protein of 355 amino acids with an estimated molecular weight of 58.0 kDa. Sequence analysis and multiple sequence alignment demonstrated that Ac-cathL resembles members of cathepsin L family of other parasites and mammals. Stage-dependent mRNA expression analysis showed that Ac-cathL transcripts were expressed in all stages of A. cantonensis, with the highest expression in female stage. The recombinant Ac-cathL (rAc-cathL) expressed in Escherichia coli exhibited protease activity in acidic pH as demonstrated by gelatin zymography, as well as hydrolytic activity against natural substrates, including BSA, human IgG and human fibrinogen. Immunolocalization revealed that Ac-cathL is localized in tegument of the 18 days post infection stage and uterus of the female adult stage. Therefore, these results implied that the Ac-cathL plays important roles in host tissue migration, nutrition uptake and immune evasion. |
| Databáze: | OpenAIRE |
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