Efficient preparation of human and mouse CD1d proteins using silkworm baculovirus expression system

Kusaka, Hiroki; Kita, Shunsuke; Tadokoro, Takashi; Yoshida, Kouki; Kasai, Yoshiyuki; Niiyama, Harumi; Fujimoto, Yukari; Hanashima, Shinya; Murata, Michio; Sugiyama, Shigeru; Ose, Toyoyuki; Kuroki, Kimiko; Maenaka, Katsumi

Efficient preparation of human and mouse CD1d proteins using silkworm baculovirus expression system

Kusaka, Hiroki; Kita, Shunsuke; Tadokoro, Takashi et al.

Protein expression and purification, 2020, 172, 105631

Permalink : https://hdl.handle.net/2115/82649

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アクセス権	open access
URI	https://hdl.handle.net/2115/82649
タイトル	Efficient preparation of human and mouse CD1d proteins using silkworm baculovirus expression system
著者	Kusaka, Hiroki ; Kita, Shunsuke ; Tadokoro, Takashi ; Yoshida, Kouki ; Kasai, Yoshiyuki ; Niiyama, Harumi ; Fujimoto, Yukari ; Hanashima, Shinya ; Murata, Michio ; Sugiyama, Shigeru ; Ose, Toyoyuki ; Kuroki, Kimiko ; Maenaka, Katsumi
著者名 Kusaka, Hiroki
著者名 Kita, Shunsuke
著者名 Tadokoro, Takashi
著者名 Yoshida, Kouki
著者名 Kasai, Yoshiyuki
著者名 Niiyama, Harumi
著者名 Fujimoto, Yukari
著者名 Hanashima, Shinya
著者名 Murata, Michio
著者名 Sugiyama, Shigeru
e-Rad_Researcher 80380525 著者名 Ose, Toyoyuki
e-Rad_Researcher 90553313 著者名 Kuroki, Kimiko
e-Rad_Researcher 10322752 著者名 Maenaka, Katsumi
言語	英語
キーワード	CD1d
	Secretory expression
	Major histocompatibility complex class I
	Differential scanning calorimetry
	Silkworm
	Baculovirus
発行日	2020-08
出版者	Elsevier
収録物名	Protein expression and purification
巻(号)	172
ページ	105631
抄録	CD1d is a major histocompatibility complex (MHC) class I-like glycoprotein and binds to glycolipid antigens that are recognized by natural killer T (NKT) cells. To date, our understanding of the structural basis for glycolipid binding and receptor recognition of CD1d is still limited. Here, we established a preparation method for the ectodomain of human and mouse CD1d using a silkworm-baculovirus expression system. The co-expression of human and mouse CD1d and beta 2-microglobulin (beta 2m) in the silkworm-baculovirus system was successful, but the yield of human CD1d was low. A construct of human CD1d fused with beta 2m via a flexible GS linker as a single polypeptide was prepared to improve protein yield. The production of this single-chained complex was higher (50 mu g/larva) than that of the co-expression complex. Furthermore, differential scanning calorimetry revealed that the linker made the CD1d complex more stable and homogenous. These results suggest that the silkworm baculovirus expression system is useful for structural and biophysical studies of CD1d in several aspects including low cost, easy handling, biohazard-free, rapid, and high yielding.
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	©2020, Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/
PISSN	1046-5928
関連情報 (isVersionOf)	出版社版DOI https://doi.org/10.1016/j.pep.2020.105631