5AE0
Perdeuterated mouse CNPase catalytic domain at atomic resolution
Summary for 5AE0
| Entry DOI | 10.2210/pdb5ae0/pdb |
| Descriptor | 2', 3'-CYCLIC-NUCLEOTIDE 3'-PHOSPHODIESTERASE, CITRIC ACID, CHLORIDE ION, ... (4 entities in total) |
| Functional Keywords | hydrolase |
| Biological source | MUS MUSCULUS (HOUSE MOUSE) |
| Total number of polymer chains | 1 |
| Total formula weight | 24521.51 |
| Authors | Laulumaa, S.,Kursula, P. (deposition date: 2015-08-25, release date: 2015-11-25, Last modification date: 2024-01-10) |
| Primary citation | 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 Cited by 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: 26563764DOI: 10.1038/srep16520 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.04 Å) |
Structure validation
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