Evolutionary upgrade of stefins for secretion in parasites

Michal Buša1, Zuzana Matoušková1, Pavlína Řezáčová1, John Dalton2,
Saša Štefanič3, Michael Mareš1

1Institute of Organic Chemistry and Biochemistry, Czech Academy of Sciences, 166 10 Prague, Czech Republic

2School of Biological Sciences, Queen's University, BT9 7BL Belfast, Northern Ireland

3Institute of Parasitology, University of Zurich, 8057 Zurich, Switzerland

mares@uochb.cas.cz

Fasciolosis caused by the liver fluke Fasciola hepatica is a worldwide spread parasitic disease of ruminants and is recognized as an emerging human disease. This work is focused on FhCY2, a member of cystatin superfamily, which is present in several tissues of F. hepatica and its excretory/secretory products. Structural and phylogenetic analyses revealed that FhCY2 belongs to stefins (type 1 cystatins). Stefins are typically intracellular, without signal sequence and without disulfides. However, FhCY2 breaks these rules, thus resembling members of extracellular type 2 cystatins. Our work shows that FhCY2 is an evolutionary adaptation to the absence of type 2 cystatins in the F. hepatica genome. FhCY2 is a broad-selective inhibitor of host cysteine cathepsins as well as secreted digestive cysteine cathepsins of F. hepatica, suggesting a dual role of FhCY2 in the physiological regulation of exogenous and endogenous proteolytic systems.