6KER

Crystal structure of D113A mutant of Drosophila melanogaster Noppera-bo, glutathione S-transferase epsilon 14 (DmGSTE14), in glutathione-bound form

これはPDB形式変換不可エントリーです。

6KER の概要

• エントリーDOI: 10.2210/pdb6ker/pdb
• 分子名称: Glutathione S-transferase E14, GLUTATHIONE (3 entities in total)
• 機能のキーワード: glutathione, glutathione s-transferase, gst, transferase
• 由来する生物種: Drosophila melanogaster (Fruit fly)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 55614.16

構造登録者

Koiwai, K.,Inaba, K.,Morohashi, K.,Yumoto, F.,Niwa, R.,Senda, T. (登録日: 2019-07-04, 公開日: 2019-10-02, 最終更新日: 2023-11-22)

主引用文献

Koiwai, K.,Inaba, K.,Morohashi, K.,Enya, S.,Arai, R.,Kojima, H.,Okabe, T.,Fujikawa, Y.,Inoue, H.,Yoshino, R.,Hirokawa, T.,Kato, K.,Fukuzawa, K.,Shimada-Niwa, Y.,Nakamura, A.,Yumoto, F.,Senda, T.,Niwa, R.
An integrated approach to unravel a crucial structural property required for the function of the insect steroidogenic Halloween protein Noppera-bo.
J.Biol.Chem., 295:7154-7167, 2020

PubMed Abstract: Ecdysteroids are the principal steroid hormones essential for insect development and physiology. In the last 18 years, several enzymes responsible for ecdysteroid biosynthesis encoded by Halloween genes were identified and genetically and biochemically characterized. However, the tertiary structures of these proteins have not yet been characterized. Here, we report the results of an integrated series of , , and analyses of the Halloween GST protein Noppera-bo (Nobo). We determined crystal structures of Nobo (DmNobo) complexed with GSH and 17β-estradiol, a DmNobo inhibitor. 17β-Estradiol almost fully occupied the putative ligand-binding pocket and a prominent hydrogen bond formed between 17β-estradiol and Asp-113 of DmNobo. We found that Asp-113 is essential for 17β-estradiol-mediated inhibition of DmNobo enzymatic activity, as 17β-estradiol did not inhibit and physically interacted less with the D113A DmNobo variant. Asp-113 is highly conserved among Nobo proteins, but not among other GSTs, implying that this residue is important for endogenous Nobo function. Indeed, a homozygous allele with the D113A substitution exhibited embryonic lethality and an undifferentiated cuticle structure, a phenocopy of complete loss-of-function homozygotes. These results suggest that the family of GST proteins has acquired a unique amino acid residue that appears to be essential for binding an endogenous sterol substrate to regulate ecdysteroid biosynthesis. To the best of our knowledge, ours is the first study describing the structural characteristics of insect steroidogenic Halloween proteins. Our findings provide insights relevant for applied entomology to develop insecticides that specifically inhibit ecdysteroid biosynthesis.

PubMed: 32241910
DOI: 10.1074/jbc.RA119.011463

実験手法

X-RAY DIFFRACTION (1.84 Å)