Molecular characterization of Histomonas meleagridis exoproteome with emphasis on protease secretion and parasite-bacteria interaction
| Autor: | Michael Hess, Rounik Mazumdar, Katharina Nöbauer, Karin Hummel, Ivana Bilic |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Proteomics
Proteome medicine.medical_treatment Protein Expression Protozoan Proteins Parabasalidea Pathology and Laboratory Medicine Biochemistry Histomonas meleagridis Poultry 0302 clinical medicine Cysteine Proteases Medicine and Health Sciences Protein Interaction Maps Enzyme Inhibitors Post-Translational Modification Protozoan Infections Animal 0303 health sciences Multidisciplinary biology Virulence Proteases Genomics Cysteine protease Enzymes Lytic cycle Medicine Pathogens Transcriptome Analysis Signal Peptides medicine.drug Research Article Virulence Factors Science 030231 tropical medicine Research and Analysis Methods Microbiology Host-Parasite Interactions 03 medical and health sciences Bacterial Proteins Exopeptidases medicine Gene Expression and Vector Techniques Genetics Animals Protease Inhibitors Shotgun proteomics Molecular Biology Techniques Molecular Biology Poultry Diseases 030304 developmental biology Molecular Biology Assays and Analysis Techniques Protease Biology and Life Sciences Proteins Computational Biology biology.organism_classification Genome Analysis Protease inhibitor (biology) Enzymology |
| Zdroj: | PLoS ONE PLoS ONE, Vol 14, Iss 2, p e0212429 (2019) |
| ISSN: | 1932-6203 |
| Popis: | The exoproteome of parasitic protists constitutes extracellular proteins that play a fundamental role in host-parasite interactions. Lytic factors, especially secreted proteases, are capable of modulating tissue invasion, thereby aggravating host susceptibility. Despite the important role of exoproteins during infection, the exoproteomic data on Histomonas meleagridis are non-existent. The present study employed traditional 1D-in-gel-zymography (1D-IGZ) and micro-LC-ESI-MS/MS (shotgun proteomics), to investigate H. meleagridis exoproteomes, obtained from a clonal virulent and an attenuated strain. Both strains were maintained as mono-eukaryotic monoxenic cultures with Escherichia coli. We demonstrated active in vitro secretion kinetics of proteases by both parasite strains, with a widespread proteolytic activity ranging from 17 kDa to 120 kDa. Based on protease inhibitor susceptibility assay, the majority of proteases present in both exoproteomes belonged to the family of cysteine proteases and showed stronger activity in the exoproteome of a virulent H. meleagridis. Shotgun proteomics, aided by customized database search, identified 176 proteins including actin, potential moonlighting glycolytic enzymes, lytic molecules such as pore-forming proteins (PFPs) and proteases like cathepsin-L like cysteine protease. To quantify the exoproteomic differences between the virulent and the attenuated H. meleagridis cultures, a sequential window acquisition of all theoretical spectra mass spectrometric (SWATH-MS) approach was applied. Surprisingly, results showed most of the exoproteomic differences to be of bacterial origin, especially targeting metabolism and locomotion. By deciphering such molecular signatures, novel insights into a complex in vitro protozoan- bacteria relationship were elucidated. |
| Databáze: | OpenAIRE |
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