Thiopurine intolerance-causing mutations in NUDT15 induce temperature-dependent destabilization of the catalytic site
Language English Country Netherlands Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
30639426
DOI
10.1016/j.bbapap.2019.01.006
PII: S1570-9639(19)30008-1
Knihovny.cz E-resources
- Keywords
- H/D exchange, NUDT15, Pharmacogenomic variants, Structural mass spectrometry, Thiopurine metabolism,
- MeSH
- Deoxyguanine Nucleotides chemistry MeSH
- Catalytic Domain MeSH
- Mutation MeSH
- Mutagenesis, Site-Directed MeSH
- Pyrophosphatases chemistry genetics MeSH
- Protein Stability MeSH
- Temperature MeSH
- Thermolysin chemistry MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Deoxyguanine Nucleotides MeSH
- deoxyguanosine triphosphate MeSH Browser
- NUDT15 protein, human MeSH Browser
- Pyrophosphatases MeSH
- Thermolysin MeSH
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.
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