Thiopurine intolerance-causing mutations in NUDT15 induce temperature-dependent destabilization of the catalytic site
| Autor: | Milan Fábry, Aleš Hnízda, Irena Sieglová, Petr Novák, Daniel Kavan, Petr Man |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Mutant Thermolysin Biophysics Context (language use) Biochemistry Analytical Chemistry 03 medical and health sciences 0302 clinical medicine Germline mutation Protein structure Catalytic Domain Pyrophosphatases Molecular Biology chemistry.chemical_classification Thiopurine methyltransferase biology Protein Stability Chemistry Temperature Deoxyguanine Nucleotides 030104 developmental biology Enzyme Protein destabilization 030220 oncology & carcinogenesis Mutation Mutagenesis Site-Directed biology.protein |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics. 1867:376-381 |
| ISSN: | 1570-9639 |
| DOI: | 10.1016/j.bbapap.2019.01.006 |
| Popis: | Germline mutations in NUDT15 cause thiopurine intolerance during treatment of leukemia or autoimmune diseases. Previously, it has been shown that the mutations affect the enzymatic activity of the NUDT15 hydrolase due to decreased protein stability in vivo. Here we provide structural insights into protein destabilization in R139C and V18I mutants using thermolysin-based proteolysis and H/D exchange followed by mass spectrometry. Both mutants exhibited destabilization of the catalytic site, which was more pronounced at higher temperature. This structural perturbation is shared by the mutations despite their different positions within the protein structure. Reaction products of NUDT15 reverted these conformational abnormalities, demonstrating the importance of ligands for stabilization of a native state of the mutants. This study shows the action of pharmacogenetic variants in NUDT15 in a context of protein structure, which might open novel directions in personalized chemotherapy. |
| Databáze: | OpenAIRE |
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