Base-Flipping Mutations of Uracil DNA Glycosylase:  Substrate Rescue Using a Pyrene Nucleotide Wedge

• Published in: Biochemistry (Easton) Vol. 41; no. 37; pp. 11248 - 11254
• Main Authors: Jiang, Yu Lin, Stivers, James T, Song, Fenhong
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 17.09.2002
• Subjects: Amino Acid Substitution - genetics; Base Pairing - genetics; DNA Glycosylases; Kinetics; Mutagenesis, Site-Directed; N-Glycosyl Hydrolases - chemistry; N-Glycosyl Hydrolases - genetics; Nucleic Acid Heteroduplexes - chemistry; Nucleic Acid Heteroduplexes - genetics; Nucleotides - chemistry; Organophosphorus Compounds - chemistry; Protein Binding - genetics; Pyrenes - chemistry; Spectrometry, Fluorescence; Substrate Specificity - genetics; Uracil - chemistry; Uracil-DNA Glycosidase

We recently introduced a new substrate rescue tool for investigating enzymatic base flipping by uracil DNA glycosylase (UDG) in which a bulky pyrene nucleotide wedge (Y) was placed opposite a uracil in duplex DNA (i.e., a U/Y pair), thereby preorganizing the target base in an extrahelical conformati...