Photoinduced Electron Transfer from the Tryptophan Triplet State in Zn-Azurin

Tryptophan is one of few residues that participates in biological electron transfer reactions. Upon substitution of the native Cu²⁺ center with Zn²⁺ in the blue-copper protein azurin, a long-lived tryptophan neutral radical can be photogenerated. We report the following quantum yield values for Zn-substituted azurin in the presence of the electron acceptor Cu(II)-azurin: formation of the tryptophan neutral radical (Φ_rad), electron transfer (Φ_ET), fluorescence (Φ_fluo), and phosphorescence (Φ_phos), as well as the efficiency of proton transfer of the cation radical (Φ_PT). Increasing the concentration of the electron acceptor increased Φ_rad and Φ_ET values and decreased Φ_phos without affecting Φ_fluo. At all concentrations of the acceptor, the value of Φ_PT was nearly unity. These observations indicate that the phosphorescent triplet state is the parent state of electron transfer and that nearly all electron transfer events lead to proton loss. Similar results regarding the parent state were obtained with a different electron acceptor, [Co(NH₃)₅Cl]²⁺; however, Stern-Volmer graphs revealed that [Co(NH₃)₅Cl]²⁺ was a more effective phosphorescence quencher (K _SV = 230 000 M^-1) compared to Cu(II)-azurin (K _SV = 88 000 M^-1). Competition experiments in the presence of both [Co(NH₃)₅Cl]²⁺ and Cu(II)-azurin suggested that [Co(NH₃)₅Cl]²⁺ is the preferred electron acceptor. Implications of these results in terms of quenching mechanisms are discussed.