Ca2+-induced structural changes and intramolecular interactions in N-terminal region of diacylglycerol kinase alpha

Daisuke Takahashi, Kento Yonezawa, Yuki Okizaki, Jose M.M. Caaveiro, Tadashi Ueda, Atsushi Shimada, Fumio Sakane, Nobutaka Shimizu

研究成果: ジャーナルへの寄稿学術誌査読

抄録

Diacylglycerol kinases (DGKs) are multi-domain lipid kinases that modulate the levels of lipid messengers, diacylglycerol, and phosphatidic acid. Recently, increasing attention has been paid to its α isozyme (DGKα) as a potential target for cancer immunotherapy. However, little progress has been made on the structural biology of DGKs, and a detailed understanding of the Ca2+-triggered activation of DGKα, for which the N-terminal domains likely play a critical role, remains unclear. We have recently shown that Ca2+ binding to DGKα-EF induces conformational changes from a protease-susceptible “open” conformation in the apo state to a well-folded one in its holo state. Here, we further studied the structural properties of DGKα N-terminal (RVH and EF) domains using a series of biophysical techniques. We first revealed that the N-terminal RVH domain is a novel Ca2+-binding domain, but the Ca2+-induced conformational changes mainly occur in the EF domain. This was corroborated by NMR experiments showing that the EF domain adopts a molten-globule like structure in the apo state. Further analyses using SEC-SAXS and NMR indicate that the partially unfolded EF domain interacts with RVH domain, likely via hydrophobic interactions in the absence of Ca2+, and this interaction is modified in the presence of Ca2+. Taken together, these results present novel insights into the structural rearrangement of DGKα N-terminal domains upon binding to Ca2+, which is essential for the activation of the enzyme.

本文言語英語
論文番号e4365
ジャーナルProtein Science
31
7
DOI
出版ステータス出版済み - 7月 2022

!!!All Science Journal Classification (ASJC) codes

  • 生化学
  • 分子生物学

フィンガープリント

「Ca2+-induced structural changes and intramolecular interactions in N-terminal region of diacylglycerol kinase alpha」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

引用スタイル