title: Neutralization of the γ-secretase activity by monoclonal antibody against extracellular domain of nicastrin.
abstract: Several lines of evidence suggest that aberrant Notch signaling contributes to the development of several types of cancer. Activation of Notch receptor is executed through intramembrane proteolysis by γ-secretase, which is a multimeric membrane-embedded protease comprised of presenilin, nicastrin (NCT), anterior pharynx defective 1 and PEN-2. In this study, we report the neutralization of the γ-secretase activity by a novel monoclonal antibody A5226A against the extracellular domain of NCT, generated by using a recombinant budded baculovirus as an immunogen. This antibody recognized fully glycosylated mature NCT in the active γ-secretase complex on the cell surface, and inhibited the γ-secretase activity by competing with the substrate binding in vitro. Moreover, A5226A abolished the γ-secretase activity-dependent growth of cancer cells in a xenograft model. Our data provide compelling evidence that NCT is a molecular target for the mechanism-based inhibition of γ-secretase, and that targeting NCT might be a novel therapeutic strategy against cancer caused by aberrant γ-secretase activity and Notch signaling.
authors: Department of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.
authors: I
authors: I
authors: Hayashi
authors: Department of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.
authors: S
authors: S
authors: Takatori
authors: Department of Systems Biology and Medicine, Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, Japan.
authors: Y
authors: Y
authors: Urano
authors: Department of Clinical and Experimental Hematology, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Japan.
authors: Y
authors: Y
authors: Miyake
authors: Laboratory of Protein Synthesis and Expression, Institute for Protein Research, Osaka University, Osaka, Japan.
authors: J
authors: J
authors: Takagi
authors: Department of Clinical and Experimental Hematology, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Japan.
authors: M
authors: M
authors: Sakata-Yanagimoto
authors: Department of Systems Biology and Medicine, Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, Japan.|Perseus Proteomics Inc., Tokyo, Japan.
authors: H
authors: H
authors: Iwanari
authors: Department of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.
authors: S
authors: S
authors: Osawa
authors: Department of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.
authors: Y
authors: Y
authors: Morohashi
authors: Department of Pathology, The Johns Hopkins University, School of Medicine, Baltimore, MD, USA.
authors: T
authors: T
authors: Li
authors: Department of Pathology, The Johns Hopkins University, School of Medicine, Baltimore, MD, USA.
authors: P C
authors: PC
authors: Wong
authors: Department of Clinical and Experimental Hematology, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Japan.
authors: S
authors: S
authors: Chiba
authors: Department of Systems Biology and Medicine, Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, Japan.
authors: T
authors: T
authors: Kodama
authors: Department of Systems Biology and Medicine, Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, Japan.
authors: T
authors: T
authors: Hamakubo
authors: Department of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.|Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, Saitama, Japan.
authors: T
authors: T
authors: Tomita
authors: Department of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.|Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, Saitama, Japan.|Department of Neuropathology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
authors: T
authors: T
authors: Iwatsubo
mesh_terms: D053829:Amyloid Precursor Protein Secretases / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D057134:Antibodies, Neutralizing / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D000918:Antibody Specificity / Q000276:immunology; D055162:Biocatalysis / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D016513:Mice, SCID; D009369:Neoplasms / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control; D009500:Neutralization Tests; D011485:Protein Binding / Q000187:drug effects; D047368:Tumor Burden / Q000187:drug effects; D023041:Xenograft Model Antitumor Assays
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D008562:Membrane Glycoproteins; C413604:nicastrin protein; D053829:Amyloid Precursor Protein Secretases