The binding property of a monoclonal antibody against the extracellular domains of aquaporin-4 directs aquaporin-4 toward endocytosis

Ping Huang, Yoshiki Takai, Osamu Kusano-Arai, Julia Ramadhanti, Hiroko Iwanari, Takayuki Miyauchi, Toshiko Sakihama, Jing Yan Han, Masashi Aoki, Takao Hamakubo, Kazuo Fujihara, Masato Yasui, Yoichiro Abe

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Neuromyelitis optica (NMO), an autoimmune disease of the central nervous system, is characterized by an autoantibody called NMO-IgG that recognizes the extracellular domains (ECDs) of aquaporin-4 (AQP4). In this study, monoclonal antibodies (mAbs) against the ECDs of mouse AQP4 were established by a baculovirus display method. Two types of mAb were obtained: one (E5415A) recognized both M1 and M23 isoforms, and the other (E5415B) almost exclusively recognized the square-array-formable M23 isoform. While E5415A enhanced endocytosis of both M1 and M23, followed by degradation in cells expressing AQP4, including astrocytes, E5415B did so to a much lesser degree, as determined by live imaging using fluorescence-labeled antibodies and by Western blotting of lysate of cells treated with these mAbs. E5415A promoted cluster formation of AQP4 on the cell surface prior to endocytosis as determined by immunofluorescent microscopic observation of bound mAbs to astrocytes as well as by Blue native PAGE analysis of AQP4 in the cells treated with the mAbs. These observations clearly indicate that an anti-AQP4-ECDs antibody possessing an ability to form a large cluster of AQP4 by cross-linking two or more tetramers outside the AQP4 arrays enhances endocytosis and the subsequent lysosomal degradation of AQP4.

Original languageEnglish
Pages (from-to)77-83
Number of pages7
JournalBiochemistry and Biophysics Reports
Volume7
DOIs
Publication statusPublished - 2016 Sep 1

Fingerprint

Aquaporin 4
Endocytosis
Monoclonal Antibodies
Neuromyelitis Optica
Astrocytes
Protein Isoforms
Autoimmune Diseases of the Nervous System
Native Polyacrylamide Gel Electrophoresis
Degradation
Antibodies
Baculoviridae
Optical Imaging
Neurology
Autoantibodies
Central Nervous System
Immunoglobulin G
Western Blotting
Fluorescence
Display devices
Cells

Keywords

  • Aquaporin-4
  • Astrocyte
  • Autoimmune disease
  • Endocytosis
  • Monoclonal antibody
  • Neuromyelitis optica

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

The binding property of a monoclonal antibody against the extracellular domains of aquaporin-4 directs aquaporin-4 toward endocytosis. / Huang, Ping; Takai, Yoshiki; Kusano-Arai, Osamu; Ramadhanti, Julia; Iwanari, Hiroko; Miyauchi, Takayuki; Sakihama, Toshiko; Han, Jing Yan; Aoki, Masashi; Hamakubo, Takao; Fujihara, Kazuo; Yasui, Masato; Abe, Yoichiro.

In: Biochemistry and Biophysics Reports, Vol. 7, 01.09.2016, p. 77-83.

Research output: Contribution to journalArticle

Huang, P, Takai, Y, Kusano-Arai, O, Ramadhanti, J, Iwanari, H, Miyauchi, T, Sakihama, T, Han, JY, Aoki, M, Hamakubo, T, Fujihara, K, Yasui, M & Abe, Y 2016, 'The binding property of a monoclonal antibody against the extracellular domains of aquaporin-4 directs aquaporin-4 toward endocytosis', Biochemistry and Biophysics Reports, vol. 7, pp. 77-83. https://doi.org/10.1016/j.bbrep.2016.05.017
Huang, Ping ; Takai, Yoshiki ; Kusano-Arai, Osamu ; Ramadhanti, Julia ; Iwanari, Hiroko ; Miyauchi, Takayuki ; Sakihama, Toshiko ; Han, Jing Yan ; Aoki, Masashi ; Hamakubo, Takao ; Fujihara, Kazuo ; Yasui, Masato ; Abe, Yoichiro. / The binding property of a monoclonal antibody against the extracellular domains of aquaporin-4 directs aquaporin-4 toward endocytosis. In: Biochemistry and Biophysics Reports. 2016 ; Vol. 7. pp. 77-83.
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