title: Cloning and Characterization of Two Potent Kunitz Type Protease Inhibitors from Echinococcus granulosus.
abstract: The tapeworm Echinococcus granulosus is responsible for cystic echinococcosis (CE), a cosmopolitan disease which imposes a significant burden on the health and economy of affected communities. Little is known about the molecular mechanisms whereby E. granulosus is able to survive in the hostile mammalian host environment, avoiding attack by host enzymes and evading immune responses, but protease inhibitors released by the parasite are likely implicated. We identified two nucleotide sequences corresponding to secreted single domain Kunitz type protease inhibitors (EgKIs) in the E. granulosus genome, and their cDNAs were cloned, bacterially expressed and purified. EgKI-1 is highly expressed in the oncosphere (egg) stage and is a potent chymotrypsin and neutrophil elastase inhibitor that binds calcium and reduced neutrophil infiltration in a local inflammation model. EgKI-2 is highly expressed in adult worms and is a potent inhibitor of trypsin. As powerful inhibitors of mammalian intestinal proteases, the EgKIs may play a pivotal protective role in preventing proteolytic enzyme attack thereby ensuring survival of E. granulosus within its mammalian hosts. EgKI-1 may also be involved in the oncosphere in host immune evasion by inhibiting neutrophil elastase and cathepsin G once this stage is exposed to the mammalian blood system. In light of their key roles in protecting E. granulosus from host enzymatic attack, the EgKI proteins represent potential intervention targets to control CE. This is important as new public health measures against CE are required, given the inefficiencies of available drugs and the current difficulties in its treatment and control. In addition, being a small sized highly potent serine protease inhibitor, and an inhibitor of neutrophil chemotaxis, EgKI-1 may have clinical potential as a novel anti-inflammatory therapeutic.
authors.affiliation: Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Herston, Queensland, Australia.|School of Public Health, The University of Queensland, Herston, Queensland, Australia.
authors.forename: Shiwanthi L
authors.initials: SL
authors.lastname: Ranasinghe
authors.affiliation: Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Herston, Queensland, Australia.
authors.forename: Katja
authors.initials: K
authors.lastname: Fischer
authors.affiliation: Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Herston, Queensland, Australia.|Department of Biochemistry, Xinjiang Medical University, Urumqi, Xinjiang, China.|Clinical Medical Research Institute, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang, China.
authors.forename: Wenbao
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Herston, Queensland, Australia.
authors.forename: Geoffrey N
authors.initials: GN
authors.lastname: Gobert
authors.affiliation: Molecular Parasitology Laboratory, QIMR Berghofer Medical Research Institute, Herston, Queensland, Australia.
authors.forename: Donald P
authors.initials: DP
authors.lastname: McManus
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D048209:Echinococcus granulosus / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D051379:Mice; D011480:Protease Inhibitors / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D011480:Protease Inhibitors; D011994:Recombinant Proteins