4WDB

Catalytic domain of mouse 2',3'-cyclic nucleotide 3'- phosphodiesterase, with mutation R307Q, complexed with 2'-AMP

4WDB の概要

• エントリーDOI: 10.2210/pdb4wdb/pdb
• 分子名称: 2',3'-cyclic-nucleotide 3'-phosphodiesterase, ADENOSINE-2'-MONOPHOSPHATE, CHLORIDE ION, ... (4 entities in total)
• 機能のキーワード: hydrolase, myelin, nervous system
• 由来する生物種: Mus musculus (House Mouse)
• 細胞内の位置: Membrane ; Lipid-anchor : P16330
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 24647.54

構造登録者

Myllykoski, M.,Raasakka, A.,Kursula, P. (登録日: 2014-09-08, 公開日: 2015-09-23, 最終更新日: 2024-01-10)

主引用文献

Raasakka, A.,Myllykoski, M.,Laulumaa, S.,Lehtimaki, M.,Hartlein, M.,Moulin, M.,Kursula, I.,Kursula, P.
Determinants of ligand binding and catalytic activity in the myelin enzyme 2',3'-cyclic nucleotide 3'-phosphodiesterase.
Sci Rep, 5:16520-16520, 2015

PubMed Abstract: 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) is an enzyme highly abundant in the central nervous system myelin of terrestrial vertebrates. The catalytic domain of CNPase belongs to the 2H phosphoesterase superfamily and catalyzes the hydrolysis of nucleoside 2',3'-cyclic monophosphates to nucleoside 2'-monophosphates. The detailed reaction mechanism and the essential catalytic amino acids involved have been described earlier, but the roles of many amino acids in the vicinity of the active site have remained unknown. Here, several CNPase catalytic domain mutants were studied using enzyme kinetics assays, thermal stability experiments, and X-ray crystallography. Additionally, the crystal structure of a perdeuterated CNPase catalytic domain was refined at atomic resolution to obtain a detailed view of the active site and the catalytic mechanism. The results specify determinants of ligand binding and novel essential residues required for CNPase catalysis. For example, the aromatic side chains of Phe235 and Tyr168 are crucial for substrate binding, and Arg307 may affect active site electrostatics and regulate loop dynamics. The β5-α7 loop, unique for CNPase in the 2H phosphoesterase family, appears to have various functions in the CNPase reaction mechanism, from coordinating the nucleophilic water molecule to providing a binding pocket for the product and being involved in product release.

PubMed: 26563764
DOI: 10.1038/srep16520

実験手法

X-RAY DIFFRACTION (1.6 Å)