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Thiopurine intolerance-causing mutations in NUDT15 induce temperature-dependent destabilization of the catalytic site
- Source :
- Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics. 1867:376-381
- Publication Year :
- 2019
- Publisher :
- Elsevier BV, 2019.
-
Abstract
- 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.
- Subjects :
- 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
Subjects
Details
- ISSN :
- 15709639
- Volume :
- 1867
- Database :
- OpenAIRE
- Journal :
- Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics
- Accession number :
- edsair.doi.dedup.....9a6ef466f70a59c58a953b57b0fb7ecf