Effect of hydrogen bonding on electronic spectra and reactivity of flavins

Riboflavin tetrabutyrate undergoes characteristic spectral changes, in both the first and second absorption band regions, upon hydrogen bonding with trichloroacetic acid of trifluoroacetic acid. On the basis of the calculated electron densities, hydrogen bonding at the heteroatoms of the isoalloxazine nucleus is considered to occur with increasing concentrations of the proton donor, first at N(1), then at O(12), O(14), and N(3)H, and finally at N(5). The idea that the major effect of the hydrogen bonding at the N(1), N(3)H, and oxygen atoms of the flavin nucleus is to facilitate the electrophilicity of the N(5) position, which was predicted by molecular orbital calculations, was supported by the observation that the hydrogen-bonded flavin in its triplet state abstracts hydrogen from the donor N-benzyl-n,n'-dimethylethylenediamine at a faster rate than do the non-hydrogen-bonded species in CCI4. The implications of the present study in the spectroscopic and catalytic properties of flavoproteins are briefly discussed.