title: Identification and characterization of the interactive proteins with cytotoxic T-lymphocyte antigen-2α.
abstract: Cytotoxic T-lymphocyte antigen-2α (CTLA-2α) is a potent inhibitor of cathepsin L-like cysteine proteases. Recombinant CTLA-2α is known to be a potent, competitive inhibitor of cathepsin L-like cysteine proteases. In this study, cathepsin L, cathepsin C, and tubulointerstitial nephritis antigen-related protein 1 (TINAGL1) were identified as novel interactive proteins of CTLA-2α by the yeast two-hybrid screening system. The direct interactions and co-localization of these proteins with CTLA-2α were confirmed using co-immunoprecipitation and immunofluorescence staining, respectively. The disulfide-bonded CTLA-2α/cathepsin L complex was isolated from mouse tissue. CTLA-2α was found to be specific and consistently expressed on the maternal side of the mouse placenta. Double immunofluorescence analysis showed that CTLA-2α was co-localized with cathepsin L, cathepsin C, and TINAGL1 in placenta. A simple cell-based fluorescence assay revealed that CTLA-2α exhibited inhibitory activity toward cathepsin C in live cells, which indicated that CTLA-2α is a novel endogenous inhibitor of cathepsin C.
authors.affiliation: a Joint Faculty of Veterinary Science , Yamaguchi University , Yamaguchi , Japan.
authors.forename: Bui Thi To
authors.initials: BT
authors.lastname: Nga
authors.forename: Claudius
authors.initials: C
authors.lastname: Luziga
authors.forename: Misa
authors.initials: M
authors.lastname: Yamamoto
authors.forename: Ken Takeshi
authors.initials: KT
authors.lastname: Kusakabe
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Yamamoto
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000943:Antigens, Differentiation / Q000235:genetics / Q000378:metabolism*; D019556:COS Cells; D020789:Cathepsin C / Q000235:genetics / Q000378:metabolism*; D056668:Cathepsin L / Q000235:genetics / Q000378:metabolism*; D002522:Chlorocebus aethiops; D004220:Disulfides / Q000737:chemistry; D005260:Female; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D054834:Lipocalins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D010920:Placenta / Q000737:chemistry / Q000378:metabolism*; D011247:Pregnancy; D011485:Protein Binding; D025941:Protein Interaction Mapping; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D015398:Signal Transduction; D020798:Two-Hybrid System Techniques
chemical_list: D000943:Antigens, Differentiation; D004220:Disulfides; D054834:Lipocalins; D009363:Neoplasm Proteins; D011994:Recombinant Proteins; C484283:Tinagl1 protein, mouse; C102076:cytotoxic T-lymphocyte antigen-2, mouse; D020789:Cathepsin C; C533919:Ctsc protein, mouse; D056668:Cathepsin L; C534278:Ctsl protein, mouse
keywords: CTLA-2α; TINAGL1; cathepsin C; cathepsin L; proteases inhibitor