Low temperature luminescence behaviours of 7-azatryptophan, 5-hydroxytryptophan and their chromophoric moieties.

The non-natural amino acids 7-azatryptophan (7AT) and 5-hydroxytryptophan (5HT) have come into significant recent prominence as novel intrinsic luminescence probes for protein structure, function and dynamics. Here, we examine the low temperature luminescence behaviours of these molecules and their respective chromophoric moieties 7-azaindole (7AI) and 5-hydroxyindole (5HI) in representative solvent media. To ascertain, in particular, the potential usefulness of 7AT and 5HT as phosphorescence probes for exploring protein environments with different hydrogen bonding characteristics, a comparison is made of the phosphorescence properties of 7AI and 5HI chromophores in frozen solutions of ethanol and ethyl acetate at 77 K. These solvent media have been chosen as representative models for polar protic and aprotic environments in proteins, respectively. Our findings indicate that one or more of the phosphorescence emission parameters (phosphorescence emission maxima, relative yield and phosphorescence lifetime) of 7AI and 5HI chromophores can serve as sensitive and discriminating probes of hydrogen bonding and related aspects of their surrounding environments. Furthermore, in a model viscous environment (glycerol at low temperatures) significant temperature dependence and red edge excitation shift (REES) effects are observed for the fluorescence emission of 7AT and its chromophoric moiety 7AI. This is consistent with pronounced dipolar relaxation properties of these molecules, and suggests interesting possibilities for exploiting REES in exploring their environmental rigidity in motionally constrained situations.