Photobehavior of merocyanine 540 bound to human serum albumin.
The photobehavior of merocyanine 540 (MC) was studied in homogeneous media (ethanol, buffer and glycerol), and in microheterogenous systems (Triton X-100 micelles and in the presence of human serum albumin) using stationary and time-resolved techniques. Merocyanine 540 in aqueous solution mostly forms aggregates, which in the presence of Triton X-100 or HSA are disaggregated. The extent of binding to HSA and its characteristics were estimated from dye absorption and fluorescence changes following protein addition; the Trp-214 fluorescence quenching was also employed to assess the extent of dye association, and physical separation was employed to evaluate the dye's apparent association constant. These results showed that dye adsorption on HSA takes place at both main protein-binding sites (I and II). This adsorption leads to dye monomerization, changing its photobehavior remarkably, with a noticeable increase in fluorescence and triplet lifetimes. These slower decays can be ascribed to a reduction of the dye photoisomerization rate. In addition, the adsorption of the dye partially protects it from the oxygen present in solution, thus reducing the apparent dye triplet-quenching rate constant. However, singlet oxygen and MC triplet quantum yields remain very low in all the systems tested. Finally, we found that the photoconsumption of merocyanine bound to HSA takes place predominantly by a type I mechanism, being more than seven times more efficient than that taking place in ethanol.
Alarcón
E
E
Department of Chemistry and Centre for Catalysis Research and Innovation, 10, Marie Curie, K1N 6N5, University of Ottawa, Ottawa, Ontario, Canada.
Aspée
A
A
González-Béjar
M
M
Edwards
A M
AM
Lissi
E
E
Scaiano
J C
JC
D000327:Adsorption; D006801:Humans; D007536:Isomerism; D017830:Octoxynol / Q000737:chemistry; D055668:Photochemical Processes; D010782:Photolysis; D017319:Photosensitizing Agents / Q000737:chemistry*; D011485:Protein Binding; D011744:Pyrimidinones / Q000737:chemistry*; D011789:Quantum Theory; D012709:Serum Albumin / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D014364:Tryptophan / Q000737:chemistry
D017319:Photosensitizing Agents; D011744:Pyrimidinones; D012709:Serum Albumin; C003954:merocyanine dye; D014364:Tryptophan; D017830:Octoxynol