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Molecular Biology Reagents> | Both full-length and truncated gray short tailed opossum Monodelphis domestica Neil2 (MdoNEIL2) sequences were synthesized and codon-optimized for expression in E. coli by GenScript and subcloned into a pET30 vector (Novagen) using NdeI and XhoI restriction sites, then transformed into BL21(DE3) plated onto LB-agar under kanamycin selection. | Get A Quote |
Oxidative damage on DNA arising from both endogenous and exogenous sources can result in base modifications that promote errors in replication as well as generate sites of base loss (abasic sites) that present unique challenges to maintaining genomic integrity. These lesions are excised by DNA glycosylases in the first step of the base excision repair pathway. Here we present the first crystal structure of a NEIL2 glycosylase, an enzyme active on cytosine oxidation products and abasic sites. The structure reveals an unusual “open” conformation not seen in NEIL1 or NEIL3 orthologs. NEIL2 is predicted to adopt a “closed” conformation when bound to its substrate. Combined crystallographic and solution scat... More