Schistosomiasis is one of a number of chronic helminth diseases of poverty that severely impact personal and societal well-being and productivity. Peptidases (proteases) are vital to successful parasitism, and can modulate host physiology and immunology. Interference of peptidase action by specific drugs or vaccines can be therapeutically beneficial. To date, research on peptidases in the schistosome parasite has focused on either the functional characterization of individual peptidases or their annotation as part of global genome or transcriptome studies. We were interested in functionally characterizing the complexity of peptidase activity operating at the host-parasite interface, therefore the excretory-secretory products of key developmental stages of Schistosoma mansoni that parasitize the human were examined. Using class specific peptidase inhibitors in combination with a multiplex substrate profiling assay, a number of unique activities derived from endo- and exo-peptidases were revealed in the excretory-secretory products of schistosomula (larval migratory worms), adults and eggs. The data highlight the complexity of the functional degradome for each developmental stage of this parasite and facilitate further enquiry to establish peptidase identity, physiological and immunological function, and utility as drug or vaccine candidates.
- MeSH
- interakce hostitele a patogenu MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- ovum metabolismus MeSH
- proteasy metabolismus sekrece MeSH
- proteiny červů metabolismus sekrece MeSH
- proteolýza MeSH
- Schistosoma mansoni růst a vývoj metabolismus fyziologie MeSH
- schistosomiasis mansoni parazitologie MeSH
- sekvence aminokyselin MeSH
- stadia vývoje MeSH
- substrátová specifita MeSH
- vazebná místa MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
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- MeSH
- aspartátové proteasy genetika sekrece izolace a purifikace MeSH
- debridement * metody MeSH
- Diptera MeSH
- genová knihovna MeSH
- helmintoterapie MeSH
- hojení ran MeSH
- hybridizace in situ MeSH
- komplementární DNA genetika MeSH
- larva * enzymologie genetika růst a vývoj MeSH
- lidé MeSH
- metaloproteasy genetika izolace a purifikace sekrece MeSH
- nekróza terapie MeSH
- proteasy * genetika izolace a purifikace sekrece MeSH
- sekvenční analýza DNA MeSH
- serinové proteasy genetika sekrece izolace a purifikace MeSH
- slinné žlázy enzymologie MeSH
- trávicí systém enzymologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
Fungal infections constitute a serious clinical problem in the group of patients receiving total parenteral nutrition. The majority of species isolated from infections of the total parenteral nutrition patients belong to Candida genus. The most important factors of Candida spp. virulence are the phenomenon of "phenotypic switching," adhesins, dimorphism of fungal cells and the secretion of hydrolytic enzymes such as proteinases and lipases, including aspartyl proteinases. We determined the proteolytic activity of yeast-like fungal strains cultured from the clinical materials of patients receiving total parenteral nutrition and detected genes encoding aspartyl proteinases in predominant species Candida glabrata--YPS2, YPS4, and YPS6, and Candida albicans--SAP1-3, SAP4, SAP5, and SAP6. C. albicans released proteinases on the various activity levels. All C. glabrata strains obtained from the clinical materials of examined and control groups exhibited secretion of the proteinases. All 13 isolates of C. albicans possessed genes SAP1-3. Gene SAP4 was detected in genome of 11 C. albicans strains, SAP5 in 6, and SAP6 in 11. Twenty-six among 31 of C. glabrata isolates contained YPS2 gene, 21 the YPS4 gene, and 28 the YPS6 gene. We observed that clinical isolates of C. albicans and C. glabrata differed in SAPs and YPSs gene profiles, respectively, and displayed differentiated proteolytic activity. We suppose that different sets of aspartyl proteinases genes as well as various proteinase-activity levels would have the influence on strains virulence.
- MeSH
- Candida albicans enzymologie genetika izolace a purifikace MeSH
- Candida glabrata enzymologie genetika izolace a purifikace MeSH
- faktory virulence genetika metabolismus MeSH
- genom fungální MeSH
- geny hub MeSH
- kandidóza mikrobiologie MeSH
- lidé MeSH
- parenterální výživa úplná škodlivé účinky MeSH
- proteasy genetika sekrece MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The predicted secretome of the opportunistic fungal pathogen Candida albicans contains more than 200 ORFs of diverse and often unknown function. Majority of the secreted proteins that have been experimentally evaluated to date are hydrolytic enzymes (proteinases, phospholipases and lipases). Acting on the interface between the pathogen and the host, the secreted hydrolases may enable invasion of host tissues, help the pathogen to avoid host defense mechanisms, or allow the microorganism to utilize host cell macromolecules as a source of nutrients. Aspartic proteinases constitute the best-characterized family of the C. albicans secreted proteins. Number of studies addressed also secreted phospholipases. Lipases and N-acetylhexosaminidase have received less attention thus far. Many ORFs that are predicted to encode secreted proteins await characterization.
Pomocou nerozpustného chromogénneho substrátu (čierny gelasfér) sa vypracovala jednoduchá, citlivá a reprodukovateľná metóda dôkazu extracelulárnych proteolytických enzýmov. Extracelu- lárne proteolytické enzýmy (produkované kalusovou alebo suspenznou kultúrou) hydrolyzujú čierny gelasfér. Zafarbenie okolo kalusovej kultúry a pod ňou je znakom činnosti kalusovou kultúrou produkovaných extracelulárnych proteolytických enzýmov. Intenzita zafarbenia kultivačného mé- dia je mierou aktivity extracelulárnych proteolytických enzýmov suspenznej kultúry.
By means of an insoluble chromogenic substrate (black gelasphere), a simple, sensitive and repro- ducible method of detection of extracellular proteolytic enzymes was worked out. Extracellular proteolytic enzymes (produced by a callus or suspension culture) hydrolyse black gelasphere. Colouring around the callus culture and under it is a sign of the activity of extracellular proteolytic enzymes produced by the callus culture. The intensity of colouring of the cultivating medium is a measure of the activity of extracellular proteolytic enzymes of the suspension culture.
- MeSH
- antitumorózní látky škodlivé účinky MeSH
- cévní endotel fyziologie účinky léků MeSH
- hemostáza fyziologie účinky léků MeSH
- koagulační faktory fyziologie MeSH
- krevní a lymfatické nemoci patofyziologie MeSH
- proteasy metabolismus sekrece MeSH
- trombocyty fyziologie účinky léků MeSH
- Publikační typ
- přehledy MeSH
