In vitro interaction and computational studies of glycosylated photosensitizers with plasma proteins

A series of glycosylated photosensitizers (porphyrin, chlorin, and isobacteriochlorin) in the presence of plasma proteins: bovine serum albumin (BSA) and human serum albumin (HSA), were investigated in a buffer at pH 7.4, using ultraviolet-visible (UV-vis) absorption and fluorescence spectroscopies. Due to the excitation of the tryptophan residue of BSA and HSA, its fluorescence emission was monitored around 340 nm. During each titration experiment and with each addition of the corresponding glycosylated photosensitizer, there was a concentration-dependent quenching of the intrinsic fluorescence of BSA and HSA. Using Stern-Volmer and double logarithmic plots we determined that fluorescence quenching was static for all molecules. We calculated the average binding constant for BSA and HSA for each porphyrin-type compound. To support our experimental studies, computational molecular docking and molecular dynamics simulations were used to identify the binding sites and binding poses of the each of the glycosylated photosensitizers onto BSA and HSA. The three compounds are binding to the Hemin site located in the subdomain IB of BSA forming strong interactions with Trp134, while they are binding to the subdomain IIA of HSA close to the Sudlow's site I, and interacting with Trp214.