4YGR

Crystal structure of HAD phosphatase from Thermococcus onnurineus

4YGR の概要

• エントリーDOI: 10.2210/pdb4ygr/pdb
• 関連するPDBエントリー: 4YGQ 4YGS
• 分子名称: Hydrolase, 2-[N-CYCLOHEXYLAMINO]ETHANE SULFONIC ACID, MAGNESIUM ION, ... (4 entities in total)
• 機能のキーワード: had phosphatase, substrate selectivity, hydrolase
• 由来する生物種: Thermococcus onnurineus (strain NA1)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 27199.59

構造登録者

Ngo, T.D.,Le, B.V.,Subramani, V.K.,Nguyen, C.M.T.,Lee, H.S.,Cho, Y.,Kim, K.K.,Hwang, H.Y. (登録日: 2015-02-26, 公開日: 2015-04-22, 最終更新日: 2024-03-20)

主引用文献

Ngo, T.D.,Le, B.V.,Subramani, V.K.,Nguyen, C.M.T.,Lee, H.S.,Cho, Y.,Kim, K.K.,Hwang, H.Y.
Structural basis for the substrate selectivity of a HAD phosphatase from Thermococcus onnurineus NA1
Biochem.Biophys.Res.Commun., 461:122-127, 2015

PubMed Abstract: Proteins in the haloalkaloic acid dehalogenase (HAD) superfamily, which is one of the largest enzyme families, is generally composed of a catalytic core domain and a cap domain. Although proteins in this family show broad substrate specificities, the mechanisms of their substrate recognition are not well understood. In this study, we identified a new substrate binding motif of HAD proteins from structural and functional analyses, and propose that this motif might be crucial for interacting with hydrophobic rings of substrates. The crystal structure of TON_0338, one of the 17 putative HAD proteins identified in a hyperthermophilic archaeon, Thermococcus onnurineus NA1, was determined as an apo-form at 2.0 Å resolution. In addition, we determined the crystal structure TON_0338 in complex with Mg(2+) or N-cyclohexyl-2-aminoethanesulfonic acid (CHES) at 1.7 Å resolution. Examination of the apo-form and CHES-bound structures revealed that CHES is sandwiched between Trp58 and Trp61, suggesting that this Trp sandwich might function as a substrate recognition motif. In the phosphatase assay, TON_0338 was shown to have high activity for flavin mononucleotide (FMN), and the docking analysis suggested that the flavin of FMN may interact with Trp58 and Trp61 in a way similar to that observed in the crystal structure. Moreover, the replacement of these tryptophan residues significantly reduced the phosphatase activity for FMN. Our results suggest that WxxW may function as a substrate binding motif in HAD proteins, and expand the diversity of their substrate recognition mode.

PubMed: 25858319
DOI: 10.1016/j.bbrc.2015.03.179

実験手法

X-RAY DIFFRACTION (1.703 Å)