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Determinants of ligand binding and catalytic activity in the myelin enzyme 2',3'-cyclic nucleotide 3'-phosphodiesterase

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Standard

Determinants of ligand binding and catalytic activity in the myelin enzyme 2',3'-cyclic nucleotide 3'-phosphodiesterase. / Raasakka, Arne; Myllykoski, Matti; Laulumaa, Saara; Lehtimäki, Mari; Härtlein, Michael; Moulin, Martine; Kursula, Inari; Kursula, Petri.

I: Scientific Reports, Bind 5, 13.11.2015, s. 16520.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

Harvard

Raasakka, A, Myllykoski, M, Laulumaa, S, Lehtimäki, M, Härtlein, M, Moulin, M, Kursula, I & Kursula, P 2015, 'Determinants of ligand binding and catalytic activity in the myelin enzyme 2',3'-cyclic nucleotide 3'-phosphodiesterase', Scientific Reports, bind 5, s. 16520. https://doi.org/10.1038/srep16520

APA

Raasakka, A., Myllykoski, M., Laulumaa, S., Lehtimäki, M., Härtlein, M., Moulin, M., Kursula, I., & Kursula, P. (2015). Determinants of ligand binding and catalytic activity in the myelin enzyme 2',3'-cyclic nucleotide 3'-phosphodiesterase. Scientific Reports, 5, 16520. https://doi.org/10.1038/srep16520

Vancouver

Raasakka A, Myllykoski M, Laulumaa S, Lehtimäki M, Härtlein M, Moulin M o.a. Determinants of ligand binding and catalytic activity in the myelin enzyme 2',3'-cyclic nucleotide 3'-phosphodiesterase. Scientific Reports. 2015 nov 13;5:16520. https://doi.org/10.1038/srep16520

Author

Raasakka, Arne ; Myllykoski, Matti ; Laulumaa, Saara ; Lehtimäki, Mari ; Härtlein, Michael ; Moulin, Martine ; Kursula, Inari ; Kursula, Petri. / Determinants of ligand binding and catalytic activity in the myelin enzyme 2',3'-cyclic nucleotide 3'-phosphodiesterase. I: Scientific Reports. 2015 ; Bind 5. s. 16520.

Bibtex

@article{697ce6d9bd2d4ca1a7ca1dd2c74984f4,
title = "Determinants of ligand binding and catalytic activity in the myelin enzyme 2',3'-cyclic nucleotide 3'-phosphodiesterase",
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. ",
keywords = "2',3'-Cyclic Nucleotide 3'-Phosphodiesterase/chemistry, Animals, Binding Sites/genetics, Biocatalysis, Catalytic Domain, Circular Dichroism, Crystallography, X-Ray, Kinetics, Ligands, Mass Spectrometry, Mice, Models, Molecular, Mutation, Myelin Sheath/metabolism, Protein Binding, Protein Stability, Protein Structure, Tertiary, Scattering, Small Angle, Static Electricity, Temperature, X-Ray Diffraction",
author = "Arne Raasakka and Matti Myllykoski and Saara Laulumaa and Mari Lehtim{\"a}ki and Michael H{\"a}rtlein and Martine Moulin and Inari Kursula and Petri Kursula",
year = "2015",
month = nov,
day = "13",
doi = "10.1038/srep16520",
language = "English",
volume = "5",
pages = "16520",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "nature publishing group",

}

RIS

TY - JOUR

T1 - Determinants of ligand binding and catalytic activity in the myelin enzyme 2',3'-cyclic nucleotide 3'-phosphodiesterase

AU - Raasakka, Arne

AU - Myllykoski, Matti

AU - Laulumaa, Saara

AU - Lehtimäki, Mari

AU - Härtlein, Michael

AU - Moulin, Martine

AU - Kursula, Inari

AU - Kursula, Petri

PY - 2015/11/13

Y1 - 2015/11/13

N2 - 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.

AB - 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.

KW - 2',3'-Cyclic Nucleotide 3'-Phosphodiesterase/chemistry

KW - Animals

KW - Binding Sites/genetics

KW - Biocatalysis

KW - Catalytic Domain

KW - Circular Dichroism

KW - Crystallography, X-Ray

KW - Kinetics

KW - Ligands

KW - Mass Spectrometry

KW - Mice

KW - Models, Molecular

KW - Mutation

KW - Myelin Sheath/metabolism

KW - Protein Binding

KW - Protein Stability

KW - Protein Structure, Tertiary

KW - Scattering, Small Angle

KW - Static Electricity

KW - Temperature

KW - X-Ray Diffraction

U2 - 10.1038/srep16520

DO - 10.1038/srep16520

M3 - Journal article

C2 - 26563764

VL - 5

SP - 16520

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

ER -

ID: 199337786