Screening for novel fluorescent nucleobase analogs (FBAs) using computational and experimental methods ‐ 2‐amino‐8‐vinylpurine (2A8VP), as a Case study

Abstract

Fluorescent nucleobase analogs (FBAs) are molecular reporters that are structurally similar to naturally occurring nucleotides but that have significantly higher emission quantum yields and red‐shifted absorption and emission maxima. These reporter molecules play an important role in the study of structure and dynamics of nucleic acids and proteins by providing deeper insight into the mechanisms of various biological processes. The experimental synthesis and characterization of FBAs usually precedes their computational characterization. Here, we start from computation to screen for fluorescent emission wavelength and fluorescence intensity thus reducing the time and material needed in developing new novel fluorescent reporters. The photophysical properties of a putative fluorescent analog 2‐amino‐8‐vinylpurine (2A8VP) have been predicted using TD‐DFT theory. Based on these results, we have attempted its synthesis starting from 2‐amino‐6‐chloro‐purine nucleobase. Several blue‐emissive molecules have been obtained, one with a particularly high quantum yield. These reaction products have been characterized by NMR, LC‐MS, and HPLC to obtain their structural, absorbance, and fluorescence properties to compare them with the computed properties of the 2A8VP target molecule.