Concomitant binding of two fluorescent probes at site-I of human serum albumin: The protein acting as a scaffold enabling fluorescence resonance energy transfer

Author(s)

Ximenes, Valdecir Farias

• Document type: Journal article
• Source: JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY B-BIOLOGY; v. 234, p. 10-pg., 2022-08-13.

Abstract

Human serum albumin (HSA) is the primary drug carrier in the blood plasma. Here, I aimed to show that two ligands can be accommodated simultaneously in the binding site-I of HSA. To do so, I studied the interaction inside the protein among site-I ligands of HSA via fluorescence resonance energy transfer (FRET), synchronous fluorescence, red edge excitation shift (REES), and induced circular dichroism (ICD). Warfarin (WAR), coumarin-153 (C153), 6-(p-toluidino)-2-naphthalenesulfonic acid sodium salt (TNS), dansylglycine (DGY), and 4-(dicya-nomethylene)-2-methyl-6-(4-dimethylaminostyryl)-4H-pyran (DCM) were enrolled in the investigation. I found that WAR can transfer energy to C153 only in the presence of the protein. In addition, the presence of WAR at site-I altered the protein microenvironment felt by C153. The alteration was detected by measuring the syn-chronous fluorescence, REES, and ICD in C153. The findings were validated by measuring the energy transfer from TNS to DCM and the alteration in synchronous fluorescence and REES. FRET was not observed using WAR as donor and DGY as acceptor. The result is consistent, as DGY is a site-II ligand at a higher WAR distance. In all studied cases, the effects were only observed in the presence of HSA. In conclusion, the protein acted as a scaffold approximating the ligands. These findings prove that more than one ligand can simultaneously be complex at site-I of HSA. (AU)

• FAPESP's process: 19/18445-5 - Synthesis, studies and applications of fluorescent and circular dichroic probes for characterization of interactions with proteins, DNA and determination of enzymatic activity
• Grantee: Valdecir Farias Ximenes
• Support Opportunities: Regular Research Grants