A new system to evaluate characteristics of Niemann-Pick C1 Like 1-mediated cholesterol transport using Xenopus laevis oocytes

Nashimoto, Shunsuke; Yagi, Saori; Takeda, Naoki; Nonaka, Miku; Takekuma, Yoh; Sugawara, Mitsuru; Sato, Yuki

A new system to evaluate characteristics of Niemann-Pick C1 Like 1-mediated cholesterol transport using Xenopus laevis oocytes

Nashimoto, Shunsuke; Yagi, Saori; Takeda, Naoki et al.

Biochimica et biophysica acta. Biomembranes, 2021, 1863(2), 183508

Permalink : https://hdl.handle.net/2115/83992

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アクセス権	open access
URI	https://hdl.handle.net/2115/83992
タイトル	A new system to evaluate characteristics of Niemann-Pick C1 Like 1-mediated cholesterol transport using Xenopus laevis oocytes
著者	Nashimoto, Shunsuke ; Yagi, Saori ; Takeda, Naoki ; Nonaka, Miku ; Takekuma, Yoh ; Sugawara, Mitsuru ; Sato, Yuki
著者名 Nashimoto, Shunsuke
著者名 Yagi, Saori
著者名 Takeda, Naoki
著者名 Nonaka, Miku
e-Rad_Researcher 00396293 著者名 Takekuma, Yoh
e-Rad_Researcher 60332467 著者名 Sugawara, Mitsuru
e-Rad_Researcher 00564981 著者名 Sato, Yuki
言語	英語
キーワード	Cholesterol
	Niemann-Pick C1-Like 1
	Ezetimibe
	Xenopus laevis oocytes
	Mixed micelle
	Emulsion
発行日	2021-02-01
出版者	Elsevier
収録物名	Biochimica et biophysica acta. Biomembranes
巻(号)	1863(2)
ページ	183508
抄録	Niemann-Pick C1 Like 1 (NPC1L1) is known to be involved in the intestinal absorption of cholesterol. For evaluating the function of NPC1L1, cell lines such as Caco-2, Madin-Darby canine kidney (MDCK) II, and McA-RH7777 have been used in previous studies, but the detailed molecular mechanism of transport has not been elucidated. In this study, the characteristics of cholesterol transport via NPC1L1 were investigated using a Xenopus laevis oocyte expression system in addition to a conventional cell line with stable expression. The transport activity of cholesterol uptake was increased in NPC1L1-overexpressed MDCK cells compared with that in mock cells, but MDCK cells expressed endogenous NPC1L1 and had high cholesterol transport activity. On the other hand, cRNA-injected oocytes expressed NPC1L1 after culturing for 5-6 days. The transport activity of cholesterol uptake was increased in NPC1L1 cRNA-injected oocytes compared with that in water-injected oocytes. In addition, the uptake of cholesterol was decreased in the presence of ezetimibe, an NPC1L1 inhibitor, in cRNA-injected oocytes but not in control oocytes, indicating that endogenous NPC1L1 is not expressed in oocytes. Furthermore, cholesterol uptake was substantially decreased in NPC1L1 L216A cRNA-injected oocytes compared with that in NPC1L1 cRNA-injected oo-cytes, indicating that leucine at position 216 of NPC1L1 is important for cholesterol transport and that an oocyte expression system is useful for mutant analysis. These results indicate that the oocyte expression system is useful for evaluating the characteristics of NPC1L1-mediated cholesterol transport and may contribute to the elucidation of the detailed molecular mechanism of cholesterol transport via NPC1L1.
資源タイプ	学術雑誌論文
出版タイプ	Accepted Manuscript
権利情報	Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
権利情報	©2021. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/
PISSN	0005-2736
関連情報 (isVersionOf)	出版社版DOI https://doi.org/10.1016/j.bbamem.2020.183508