Language
English
Publication Date
1-1-2024
Journal
Biotechniques
DOI
10.1080/07366205.2024.2352277
PMID
39185783
Abstract
O6-Methylguanine-DNA methyltransferase (MGMT) is a DNA repair enzyme that is overexpressed in certain tumors and is associated with resistance to the DNA alkylating agent temozolomide. MGMT inhibitors show potential in combating temozolomide resistance, but current assays for MGMT enzyme activity and inhibition, primarily oligonucleotide-based and fluorescent probe-based, are laborious and costly. The clinical relevance of temozolomide therapy calls for more convenient methodologies to study MGMT inhibition. Here, we extended the application of SNAP-Capture magnetic beads to develop a novel MGMT inhibition assay that demonstrated efficacy not only with known MGMT inhibitors, but also with the aldehyde dehydrogenase inhibitor, disulfiram. The assay uses standard fluorescence microscopy as a simple and reliable detection method, and is translationally applicable in drug discovery programs.
Keywords
Humans, O(6)-Methylguanine-DNA Methyltransferase, Enzyme Inhibitors, Disulfiram, Temozolomide, Microscopy, Fluorescence, GFP, Lomeguatrib, MGMT inhibitor, O6-benzylguanine, O6-methylguanine methyl transferase, SNAP-tag, disulfiram, temozolomide
Published Open-Access
yes
Recommended Citation
Khalaj, Vahid; AghaAmiri, Solmaz; Ghosh, Sukhen C; et al., "A Simple and Rapid In Vitro Assay for Identification of Direct Inhibitors of O6-Methylguanine-DNA Methyltransferase" (2024). The Brown Foundation: Institute of Molecular Medicine. 12.
https://digitalcommons.library.tmc.edu/molecular_med/12