7D2O

Solution structure of Gaussia Luciferase by NMR

7D2O の概要

• エントリーDOI: 10.2210/pdb7d2o/pdb
• NMR情報: BMRB: 36385
• 分子名称: Luciferase (1 entity in total)
• 機能のキーワード: luciferase, luminescent protein
• 由来する生物種: Gaussia princeps (T. Scott, 1894)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 18851.84

構造登録者

Kobayashi, N.,Wu, N.,Kuroda, Y.,Yamazaki, T. (登録日: 2020-09-17, 公開日: 2020-12-02, 最終更新日: 2024-11-13)

主引用文献

Wu, N.,Kobayashi, N.,Tsuda, K.,Unzai, S.,Saotome, T.,Kuroda, Y.,Yamazaki, T.
Solution structure of Gaussia Luciferase with five disulfide bonds and identification of a putative coelenterazine binding cavity by heteronuclear NMR.
Sci Rep, 10:20069-20069, 2020

PubMed Abstract: Gaussia luciferase (GLuc) is a small luciferase (18.2 kDa; 168 residues) and is thus attracting much attention as a reporter protein, but the lack of structural information is hampering further application. Here, we report the first solution structure of a fully active, recombinant GLuc determined by heteronuclear multidimensional NMR. We obtained a natively folded GLuc by bacterial expression and efficient refolding using a Solubility Enhancement Petide (SEP) tag. Almost perfect assignments of GLuc's H, C and N backbone signals were obtained. GLuc structure was determined using CYANA, which automatically identified over 2500 NOEs of which > 570 were long-range. GLuc is an all-alpha-helix protein made of nine helices. The region spanning residues 10-18, 36-81, 96-145 and containing eight out of the nine helices was determined with a C-atom RMSD of 1.39 Å ± 0.39 Å. The structure of GLuc is novel and unique. Two homologous sequential repeats form two anti-parallel bundles made by 4 helices and tied together by three disulfide bonds. The N-terminal helix 1 is grabbed by these 4 helices. Further, we found a hydrophobic cavity where several residues responsible for bioluminescence were identified in previous mutational studies, and we thus hypothesize that this is a catalytic cavity, where the hydrophobic coelenterazine binds and the bioluminescence reaction takes place.

PubMed: 33208800
DOI: 10.1038/s41598-020-76486-4

実験手法

SOLUTION NMR