4R8S

Dengue serotype 3 methyltransferase bound to Sinefungin

4R8S の概要

• エントリーDOI: 10.2210/pdb4r8s/pdb
• 関連するPDBエントリー: 4R8R
• 分子名称: nonstructural protein NS5, SINEFUNGIN (3 entities in total)
• 機能のキーワード: n7 methyltransferase, 2'o methyltransferase, transferase
• 由来する生物種: Dengue virus 3
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 60901.95

構造登録者

Noble, C.G. (登録日: 2014-09-02, 公開日: 2014-10-15, 最終更新日: 2023-11-08)

主引用文献

Noble, C.G.,Li, S.H.,Dong, H.,Chew, S.H.,Shi, P.Y.
Crystal structure of dengue virus methyltransferase without S-adenosyl-L-methionine
Antiviral Res., 111C:78-81, 2014

PubMed Abstract: Flavivirus methyltransferase is a genetically-validated antiviral target. Crystal structures of almost all available flavivirus methyltransferases contain S-adenosyl-L-methionine (SAM), the methyl donor molecule that co-purifies with the enzymes. This raises a possibility that SAM is an integral structural component required for the folding of dengue virus (DENV) methyltransferase. Here we exclude this possibility by solving the crystal structure of DENV methyltransferase without SAM. The SAM ligand was removed from the enzyme through a urea-mediated denaturation-and-renaturation protocol. The crystal structure of the SAM-depleted enzyme exhibits a vacant SAM-binding pocket, with a conformation identical to that of the SAM-enzyme co-crystal structure. Functionally, equivalent enzymatic activities (N-7 methylation, 2'-O methylation, and GMP-enzyme complex formation) were detected for the SAM-depleted and SAM-containing recombinant proteins. These results clearly indicate that the SAM molecule is not an essential component for the correct folding of DENV methyltransferase. Furthermore, the results imply a potential antiviral approach to search for inhibitors that can bind to the SAM-binding pocket and compete against SAM binding. To demonstrate this potential, we have soaked crystals of DENV methyltransferase without a bound SAM with the natural product Sinefungin and show that preformed crystals are capable of binding ligands in this pocket.

PubMed: 25241250
DOI: 10.1016/j.antiviral.2014.09.003

実験手法

X-RAY DIFFRACTION (1.48 Å)