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

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

Research output: Contribution to journalArticle

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 β2-microglobulin (β2m) in the silkworm-baculovirus system was successful, but the yield of human CD1d was low. A construct of human CD1d fused with β2m 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 μg/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.

Original languageEnglish
Article number105631
JournalProtein Expression and Purification
Volume172
DOIs
Publication statusPublished - 2020 Aug

Keywords

  • Baculovirus
  • CD1d
  • Differential scanning calorimetry
  • Major histocompatibility complex class I
  • Secretory expression
  • Silkworm

ASJC Scopus subject areas

  • Biotechnology

Fingerprint Dive into the research topics of 'Efficient preparation of human and mouse CD1d proteins using silkworm baculovirus expression system'. Together they form a unique fingerprint.

  • Cite this

    Kusaka, H., Kita, S., Tadokoro, T., Yoshida, K., Kasai, Y., Niiyama, H., Fujimoto, Y., Hanashima, S., Murata, M., Sugiyama, S., Ose, T., Kuroki, K., & Maenaka, K. (2020). Efficient preparation of human and mouse CD1d proteins using silkworm baculovirus expression system. Protein Expression and Purification, 172, [105631]. https://doi.org/10.1016/j.pep.2020.105631