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• W2554088269 endingPage "12248" @default.
• W2554088269 startingPage "12232" @default.
• W2554088269 abstract "The structure and dynamic motions of bases in DNA duplexes and other constructs are important for understanding mechanisms of selectivity and recognition of DNA-binding proteins. The fluorescent guanine analogue, 6-methylisoxanthopterin 6-MI, is well suited to this purpose as it exhibits an unexpected 3- to 4-fold increase in relative quantum yield upon duplex formation when incorporated into the following sequences: ATFAA, AAFTA, or ATFTA (where F represents 6-MI). To better understand some of the factors leading to the 6-MI fluorescence increase upon duplex formation, we characterized the effect of local sequence and structural perturbations on 6-MI photophysics through temperature melts, quantum yield measurements, fluorescence quenching assays, and fluorescence lifetime measurements. By examining 21 sequences we have determined that the duplex-enhanced fluorescence (DEF) depends on the composition of bases adjacent to 6-MI and the presence of adenines at locations n ± 2 from the probe. Investigation of duplex stability and local solvent accessibility measurements support a model in which the DEF arises from a constrained geometry of 6-MI in the duplex, which remains H-bonded to cytosine, stacked with adjacent bases and inaccessible to quenchers. Perturbation of DNA structure through the introduction of an unpaired base 3' to 6-MI or a mismatched basepair increases 6-MI dynamic motion leading to fluorescence quenching and a reduction in quantum yield. Molecular dynamics simulations suggest the enhanced fluorescence results from a greater degree of twist at the X-F step relative to the quenched duplexes examined. These results point to a model where adenine residues located at n ± 2 from 6-MI induce a structural geometry with greater twist in the duplex that hinders local motion reducing dynamic quenching and producing an increase in 6-MI fluorescence." @default.
• W2554088269 created "2016-11-30" @default.
• W2554088269 creator A5002606961 @default.
• W2554088269 creator A5012217609 @default.
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• W2554088269 date "2016-11-23" @default.
• W2554088269 modified "2023-09-26" @default.
• W2554088269 title "Photophysical Characterization of Enhanced 6-Methylisoxanthopterin Fluorescence in Duplex DNA" @default.
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• W2554088269 doi "https://doi.org/10.1021/acs.jpcb.6b07369" @default.
• W2554088269 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/27934220" @default.
• W2554088269 hasPublicationYear "2016" @default.
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