Functional expression and characterization of cathepsin B and L from the gut of the tick Ixodes ricinus

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60077344%3A_____%2F09%3A00336472" target="_blank" >RIV/60077344:_____/09:00336472 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/61388963:_____/09:00336472

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Functional expression and characterization of cathepsin B and L from the gut of the tick Ixodes ricinus

Popis výsledku v původním jazyce

Ticks differ from hematophagous insects in intracellular localization of hemoglobin proteolysis. We have recently described in the hard tick Ixodes ricinus that this process occurs at acidic pH and as is maintained by a machinery of gut-associated cysteine and aspartic proteases similar to blood feeding platyhelmints and nematodes. In this work, we functionally expressed in Pichia pastoris and characterized two components of tick hemoglobinolytic cascade, namely IrCB and IrCL (cysteine proteases of CA family). Using a positional scanning of synthetic combinatorial library we identified the P1?P4 specifities of both enzymes. IrCL displays a strict acidic optimum at pH 4, while IrCB shows broader pH range. Both fusion enzymes were demonstrated to digestbovine hemoglobin and albumin in vitro. Expression profiles of both enzymes were strongly upregulated during blood meal uptake.

Název v anglickém jazyce

Functional expression and characterization of cathepsin B and L from the gut of the tick Ixodes ricinus

Popis výsledku anglicky

Ticks differ from hematophagous insects in intracellular localization of hemoglobin proteolysis. We have recently described in the hard tick Ixodes ricinus that this process occurs at acidic pH and as is maintained by a machinery of gut-associated cysteine and aspartic proteases similar to blood feeding platyhelmints and nematodes. In this work, we functionally expressed in Pichia pastoris and characterized two components of tick hemoglobinolytic cascade, namely IrCB and IrCL (cysteine proteases of CA family). Using a positional scanning of synthetic combinatorial library we identified the P1?P4 specifities of both enzymes. IrCL displays a strict acidic optimum at pH 4, while IrCB shows broader pH range. Both fusion enzymes were demonstrated to digestbovine hemoglobin and albumin in vitro. Expression profiles of both enzymes were strongly upregulated during blood meal uptake.

Druh

O - Ostatní výsledky

CEP obor

EB - Genetika a molekulární biologie

OECD FORD obor

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Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>Z - Vyzkumny zamer (s odkazem do CEZ)

Rok uplatnění

2009

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů