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Title: Ultra-high field NMR studies of antibody binding and site-specific phosphorylation of {alpha}-synuclein

Abstract

Although biological importance of intrinsically disordered proteins is becoming recognized, NMR analyses of this class of proteins remain as tasks with more challenge because of poor chemical shift dispersion. It is expected that ultra-high field NMR spectroscopy offers improved resolution to cope with this difficulty. Here, we report an ultra-high field NMR study of {alpha}-synuclein, an intrinsically disordered protein identified as the major component of the Lewy bodies. Based on NMR spectral data collected at a 920 MHz proton frequency, we performed epitope mapping of an anti-{alpha}-synuclein monoclonal antibody, and furthermore, characterized conformational effects of phosphorylation at Ser129 of {alpha}-synuclein.

Authors:
 [1]; ;  [2];  [3]; ;  [2];  [4];  [5];  [6];  [7];  [8]
  1. Department of Structural Biology and Biomolecular Engineering, Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori, Mizuho-ku, Nagoya 467-8603 (Japan)|[Institute for Molecular Science, National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787 (Japan)
  2. Dept. of Structural Biology and Biomolecular Engineering, Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori, Mizuho-ku, Nagoya 467-8603 (Japan)
  3. Department of Molecular Neurobiology, Tokyo Institute of Psychiatry, 2-1-8 Kamikitazawa, Setagaya-ku, Tokyo 156-8585 (Japan)|[Molecular Neuroscience Lab., Graduate School of Science, Tokyo Metropolitan University, 1-1 Minami-Osawa, Hachioji-shi, Tokyo 192-0397 (Japan)
  4. Bioscience Research Lab., Fujiya Co., Ltd., Hadano, Kanagawa 257-0031 (Japan)
  5. Molecular Neuroscience Lab., Graduate School of Science, Tokyo Metropolitan University, 1-1 Minami-Osawa, Hachioji-shi, Tokyo 192-0397 (Japan)
  6. Dept. of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033 (Japan)
  7. Dept. of Molecular Neurobiology, Tokyo Institute of Psychiatry, 2-1-8 Kamikitazawa, Setagaya-ku, Tokyo 156-8585 (Japan)
  8. Dept. of Structural Biology and Biomolecular Engineering, Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori, Mizuho-ku, Nagoya 467-8603 (Japan)|[Institute for Molecular Science, National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787 (Japan), E-mail: kkato@phar.nagoya-cu.ac.jp
Publication Date:
OSTI Identifier:
21032987
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 363; Journal Issue: 3; Other Information: DOI: 10.1016/j.bbrc.2007.09.048; PII: S0006-291X(07)02007-4; Copyright (c) 2007 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; CASEIN; CHEMICAL SHIFT; MONOCLONAL ANTIBODIES; NUCLEAR MAGNETIC RESONANCE; PHOSPHORYLATION; PROTONS; SPECTROSCOPY

Citation Formats

Sasakawa, Hiroaki, Sakata, Eri, Yamaguchi, Yoshiki, Masuda, Masami, Mori, Tetsuya, Kurimoto, Eiji, Iguchi, Takeshi, Hisanaga, Shin-ichi, Iwatsubo, Takeshi, Hasegawa, Masato, and Kato, Koichi. Ultra-high field NMR studies of antibody binding and site-specific phosphorylation of {alpha}-synuclein. United States: N. p., 2007. Web. doi:10.1016/j.bbrc.2007.09.048.
Sasakawa, Hiroaki, Sakata, Eri, Yamaguchi, Yoshiki, Masuda, Masami, Mori, Tetsuya, Kurimoto, Eiji, Iguchi, Takeshi, Hisanaga, Shin-ichi, Iwatsubo, Takeshi, Hasegawa, Masato, & Kato, Koichi. Ultra-high field NMR studies of antibody binding and site-specific phosphorylation of {alpha}-synuclein. United States. doi:10.1016/j.bbrc.2007.09.048.
Sasakawa, Hiroaki, Sakata, Eri, Yamaguchi, Yoshiki, Masuda, Masami, Mori, Tetsuya, Kurimoto, Eiji, Iguchi, Takeshi, Hisanaga, Shin-ichi, Iwatsubo, Takeshi, Hasegawa, Masato, and Kato, Koichi. Fri . "Ultra-high field NMR studies of antibody binding and site-specific phosphorylation of {alpha}-synuclein". United States. doi:10.1016/j.bbrc.2007.09.048.
@article{osti_21032987,
title = {Ultra-high field NMR studies of antibody binding and site-specific phosphorylation of {alpha}-synuclein},
author = {Sasakawa, Hiroaki and Sakata, Eri and Yamaguchi, Yoshiki and Masuda, Masami and Mori, Tetsuya and Kurimoto, Eiji and Iguchi, Takeshi and Hisanaga, Shin-ichi and Iwatsubo, Takeshi and Hasegawa, Masato and Kato, Koichi},
abstractNote = {Although biological importance of intrinsically disordered proteins is becoming recognized, NMR analyses of this class of proteins remain as tasks with more challenge because of poor chemical shift dispersion. It is expected that ultra-high field NMR spectroscopy offers improved resolution to cope with this difficulty. Here, we report an ultra-high field NMR study of {alpha}-synuclein, an intrinsically disordered protein identified as the major component of the Lewy bodies. Based on NMR spectral data collected at a 920 MHz proton frequency, we performed epitope mapping of an anti-{alpha}-synuclein monoclonal antibody, and furthermore, characterized conformational effects of phosphorylation at Ser129 of {alpha}-synuclein.},
doi = {10.1016/j.bbrc.2007.09.048},
journal = {Biochemical and Biophysical Research Communications},
number = 3,
volume = 363,
place = {United States},
year = {Fri Nov 23 00:00:00 EST 2007},
month = {Fri Nov 23 00:00:00 EST 2007}
}
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