A theoretical analysis of the potential role of π-π stacking interactions in the photoprotolytic cycle of firefly luciferin.

Firefly oxyluciferin is a photoacid that presents a pH-sensitive fluorescence, which results from pH-dependent changes on the conformation of self-aggregated π-π stacking complexes. Luciferin is a derivative of oxyluciferin with very similar fluorescence and photoacidic properties. This similarity indicates that luciferin is also expected to be able to form π-π stacking complexes, but no pH-sensitive fluorescence is found for this compound. Here, a theoretical approach is used to rationalize this finding. We have found that luciferin only forms π-π stacking complexes in the ground state at acidic pH. At basic pH and in the excited state, luciferin is present as a dianion. This species is not able to self-aggregate, owing to repulsive electrostatic interactions. Thus, this emissive species is not subject to π-π stacking interactions; this explains its pH-insensitive fluorescence.