| Title: | Efficient preparation of human and mouse CD1d proteins using silkworm baculovirus expression system |
| Authors: | Kusaka, Hiroki Browse this author |
| Kita, Shunsuke Browse this author |
| Tadokoro, Takashi Browse this author |
| Yoshida, Kouki Browse this author |
| Kasai, Yoshiyuki Browse this author |
| Niiyama, Harumi Browse this author |
| Fujimoto, Yukari Browse this author |
| Hanashima, Shinya Browse this author |
| Murata, Michio Browse this author |
| Sugiyama, Shigeru Browse this author |
| Ose, Toyoyuki Browse this author →KAKEN DB |
| Kuroki, Kimiko Browse this author →KAKEN DB |
| Maenaka, Katsumi Browse this author →KAKEN DB |
| Keywords: | CD1d |
| Secretory expression |
| Major histocompatibility complex class I |
| Differential scanning calorimetry |
| Silkworm |
| Baculovirus |
| Issue Date: | Aug-2020 |
| Publisher: | Elsevier |
| Journal Title: | Protein expression and purification |
| Volume: | 172 |
| Start Page: | 105631 |
| Publisher DOI: | 10.1016/j.pep.2020.105631 |
| Abstract: | 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. |
| Rights: | ©2020, Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ |
| http://creativecommons.org/licenses/by-nc-nd/4.0/ |
| Type: | article (author version) |
| URI: | http://hdl.handle.net/2115/82649 |
| Appears in Collections: | 薬学研究院 (Faculty of Pharmaceutical Sciences) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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