ID | 112359 |
著者 |
Kimura, Hitoshi
Kyoto Pharmaceutical University|Tokushima University
Mikawa, Shiho
Kyoto Pharmaceutical University|Tokushima University
Mizuguchi, Chiharu
Kyoto Pharmaceutical University|Tokushima University
Horie, Yuki
Kobe Pharmaceutical University
Morita, Izumi
Kobe Pharmaceutical University
Oyama, Hiroyuki
Kobe Pharmaceutical University
Ohgita, Takashi
Kyoto Pharmaceutical University
Takeuchi, Atsuko
Kobe Pharmaceutical University
Lund-Katz, Sissel
University of Pennsylvania
Akaji, Kenichi
Kyoto Pharmaceutical University
Kobayashi, Norihiro
Kobe Pharmaceutical University
|
資料タイプ |
学術雑誌論文
|
抄録 | Apolipoprotein A-I (apoA-I) undergoes a large conformational reorganization during remodeling of high-density lipoprotein (HDL) particles. To detect structural transition of apoA-I upon HDL formation, we developed novel monoclonal antibodies (mAbs). Splenocytes from BALB/c mice immunized with a recombinant human apoA-I, with or without conjugation with keyhole limpet hemocyanin, were fused with P3/NS1/1-Ag4-1 myeloma cells. After the HAT-selection and cloning, we established nine hybridoma clones secreting anti-apoA-I mAbs in which four mAbs recognize epitopes on the N-terminal half of apoA-I while the other five mAbs recognize the central region. ELISA and bio-layer interferometry measurements demonstrated that mAbs whose epitopes are within residues 1–43 or 44–65 obviously discriminate discoidal and spherical reconstituted HDL particles despite their great reactivities to lipid-free apoA-I and plasma HDL, suggesting the possibility of these mAbs to detect structural transition of apoA-I on HDL. Importantly, a helix-disrupting mutation of W50R into residues 44–65 restored the immunoreactivity of mAbs whose epitope being within residues 44–65 against reconstituted HDL particles, indicating that these mAbs specifically recognize the epitope region in a random coil state. These results encourage us to develop mAbs targeting epitopes in the N-terminal residues of apoA-I as useful probes for monitoring formation and remodeling of HDL particles.
|
掲載誌名 |
Scientific Reports
|
ISSN | 20452322
|
出版者 | Springer Nature
|
巻 | 7
|
開始ページ | 2988
|
発行日 | 2017-06-07
|
備考 | Supplementary information : srep_7_2988_s1.pdf
|
権利情報 | © The Author(s) 2017
This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
EDB ID | |
出版社版DOI | |
出版社版URL | |
フルテキストファイル | |
言語 |
eng
|
著者版フラグ |
出版社版
|
部局 |
医学系
薬学系
|