Neutralization of the γ-secretase activity by monoclonal antibody against extracellular domain of nicastrin

I. Hayashi; S. Takatori; Y. Urano; Y. Miyake; J. Takagi; M. Sakata-Yanagimoto; H. Iwanari; S. Osawa; Y. Morohashi; T. Li; P. C. Wong; S. Chiba; T. Kodama; T. Hamakubo; T. Tomita; T. Iwatsubo

doi:10.1038/onc.2011.265

Neutralization of the γ-secretase activity by monoclonal antibody against extracellular domain of nicastrin

I. Hayashi, S. Takatori, Y. Urano, Y. Miyake, J. Takagi, M. Sakata-Yanagimoto, H. Iwanari, S. Osawa, Y. Morohashi, T. Li, P. C. Wong, S. Chiba, T. Kodama, T. Hamakubo, T. Tomita, T. Iwatsubo

School of Medicine

Research output: Contribution to journal › Article › peer-review

57 Scopus citations

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.

Original language	English (US)
Pages (from-to)	787-798
Number of pages	12
Journal	Oncogene
Volume	31
Issue number	6
DOIs	https://doi.org/10.1038/onc.2011.265
State	Published - Feb 2012

Keywords

Notch
functional antibody
g-secretase
intramembrane cleavage
membrane protein

ASJC Scopus subject areas

Molecular Biology
Genetics
Cancer Research

Access to Document

10.1038/onc.2011.265

Cite this

Hayashi, I., Takatori, S., Urano, Y., Miyake, Y., Takagi, J., Sakata-Yanagimoto, M., Iwanari, H., Osawa, S., Morohashi, Y., Li, T., Wong, P. C., Chiba, S., Kodama, T., Hamakubo, T., Tomita, T., & Iwatsubo, T. (2012). Neutralization of the γ-secretase activity by monoclonal antibody against extracellular domain of nicastrin. Oncogene, 31(6), 787-798. https://doi.org/10.1038/onc.2011.265

Hayashi, I, Takatori, S, Urano, Y, Miyake, Y, Takagi, J, Sakata-Yanagimoto, M, Iwanari, H, Osawa, S, Morohashi, Y, Li, T, Wong, PC, Chiba, S, Kodama, T, Hamakubo, T, Tomita, T & Iwatsubo, T 2012, 'Neutralization of the γ-secretase activity by monoclonal antibody against extracellular domain of nicastrin', Oncogene, vol. 31, no. 6, pp. 787-798. https://doi.org/10.1038/onc.2011.265

@article{2758c95954ac4682b33f6d802bc11a81,

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.",

keywords = "Notch, functional antibody, g-secretase, intramembrane cleavage, membrane protein",

author = "I. Hayashi and S. Takatori and Y. Urano and Y. Miyake and J. Takagi and M. Sakata-Yanagimoto and H. Iwanari and S. Osawa and Y. Morohashi and T. Li and Wong, {P. C.} and S. Chiba and T. Kodama and T. Hamakubo and T. Tomita and T. Iwatsubo",

note = "Funding Information: Southwestern Medical Center) for valuable reagents and our current and previous laboratory members for helpful discussions. We also would like to thank Keiko Tamura-Kawakami and Maiko Nampo for their excellent technical support. This work is supported in part by Grants-in-Aid for Young Scientists (S) (for TT) from Japan Society for the Promotion of Science (JSPS), by the Program for Promotion of Fundamental Studies in Health Sciences of the National",

year = "2012",

month = feb,

doi = "10.1038/onc.2011.265",

language = "English (US)",

volume = "31",

pages = "787--798",

journal = "Oncogene",

issn = "0950-9232",

publisher = "Nature Publishing Group",

number = "6",

}

TY - JOUR

T1 - Neutralization of the γ-secretase activity by monoclonal antibody against extracellular domain of nicastrin

AU - Hayashi, I.

AU - Takatori, S.

AU - Urano, Y.

AU - Miyake, Y.

AU - Takagi, J.

AU - Sakata-Yanagimoto, M.

AU - Iwanari, H.

AU - Osawa, S.

AU - Morohashi, Y.

AU - Li, T.

AU - Wong, P. C.

AU - Chiba, S.

AU - Kodama, T.

AU - Hamakubo, T.

AU - Tomita, T.

AU - Iwatsubo, T.

N1 - Funding Information: Southwestern Medical Center) for valuable reagents and our current and previous laboratory members for helpful discussions. We also would like to thank Keiko Tamura-Kawakami and Maiko Nampo for their excellent technical support. This work is supported in part by Grants-in-Aid for Young Scientists (S) (for TT) from Japan Society for the Promotion of Science (JSPS), by the Program for Promotion of Fundamental Studies in Health Sciences of the National

PY - 2012/2

Y1 - 2012/2

N2 - 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.

AB - 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.

KW - Notch

KW - functional antibody

KW - g-secretase

KW - intramembrane cleavage

KW - membrane protein

UR - http://www.scopus.com/inward/record.url?scp=84856966635&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84856966635&partnerID=8YFLogxK

U2 - 10.1038/onc.2011.265

DO - 10.1038/onc.2011.265

M3 - Article

C2 - 21725355

AN - SCOPUS:84856966635

SN - 0950-9232

VL - 31

SP - 787

EP - 798

JO - Oncogene

JF - Oncogene

IS - 6

ER -

Neutralization of the γ-secretase activity by monoclonal antibody against extracellular domain of nicastrin

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this