title: Echinococcus multilocularis: identification and functional characterization of cathepsin B-like peptidases from metacestode.
abstract: Cysteine peptidases have potent activities in the pathogenesis of various parasitic infections, and are considered as targets for chemotherapy and antigens for vaccine. In this study, two cathepsin B-like cysteine peptidases (EmCBP1 and EmCBP2) from Echinococcus multilocularis metacestodes were identified and characterized. Immunoblot analyses demonstrated that EmCBP1 and EmCBP2 were present in excretory/secretory products and extracts of E. multilocularis metacestodes. By immunohistochemistry, EmCBP1 and EmCBP2 were shown to localize to the germinal layer, the brood capsule and the protoscolex. Recombinant EmCBP1 and EmCBP2 expressed in Pichia pastoris, at optimum pH 5.5, exhibited substrate preferences for Z-Phe-Arg-MCA, Z-Val-Val-Arg-MCA, and Z-Leu-Arg-MCA, and low levels of hydrolysis of Z-Arg-Arg-MCA. Furthermore, recombinant enzymes degraded IgG, albumin, type I and IV collagens, and fibronectin. These results suggested that EmCBP1 and EmCBP2 may play key roles in protein digestion for parasites' nutrition and in parasite-host interactions.
authors.affiliation: Department of Parasitology, Asahikawa Medical University, Midorigaoka Higashi 2-1, Asahikawa, 078-8510 Hokkaido, Japan. yasusako@asahikawa-med.ac.jp
authors.forename: Yasuhito
authors.initials: Y
authors.lastname: Sako
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Nakaya
authors.forename: Akira
authors.initials: A
authors.lastname: Ito
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002401:Cathepsin B / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D004443:Echinococcosis / Q000469:parasitology*; D048210:Echinococcus multilocularis / Q000201:enzymology* / Q000235:genetics / Q000276:immunology; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D006790:Host-Parasite Interactions; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D013379:Substrate Specificity
chemical_list: D002401:Cathepsin B