title: Comparison of the binding behavior of FCCP with HSA and HTF as determined by spectroscopic and molecular modeling techniques.
abstract: The interaction of carbonylcyanide p-(trifluoromethoxy) phenylhydrazone (FCCP) with human serum albumin (HSA) and human transferrin (HTF) was investigated using multiple spectroscopy, molecular modeling, zeta-potential and conductometry measurements of aqueous solutions at pH 7.4. The fluorescence, UV/vis and polarization fluorescence spectroscopy data disclosed that the drug-protein complex formation occurred through a remarkable static quenching. Based on the fluorescence quenching, two sets of binding sites with distinct affinities for FCCP existed in the two proteins. Steady-state and polarization fluorescence analysis showed that there were more affinities between FCCP and HSA than HTF. Far UV-CD and synchronous fluorescence studies indicated that FCCP induced more structural changes on HSA. The resonance light scattering (RLS) and zeta-potential measurements suggested that HTF had a greater resistance to drug aggregation, whereas conductometry measurements expressed the presence of free ions improving the resistance of HSA to aggregation. Thermodynamic measurements implied that a combination of electrostatic and hydrophobic forces was involved in the interaction between FCCP with both proteins. The phase diagram plots indicated that the presence of second binding site on HSA and HTF was due to the existence of intermediate structures. Site marker competitive experiments demonstrated that FCCP had two distinct binding sites in HSA which were located in sub-domains IIA and IIIA and one binding site in the C-lobe of HTF as confirmed by molecular modeling. The obtained results suggested that both proteins could act as drug carriers, but that the HSA potentially had a higher capacity for delivering FCCP to cancerous tissues.
authors.affiliation: Department of Biology, Faculty of Sciences, Mashhad Branch, Islamic Azad University, Mashhad, Iran.
authors.forename: Maryam Mahmoodian
authors.initials: MM
authors.lastname: Moghaddam
authors.forename: Malihe
authors.initials: M
authors.lastname: Pirouzi
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Saberi
authors.forename: Jamshidkhan
authors.initials: J
authors.lastname: Chamani
mesh_terms: D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000737:chemistry*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007700:Kinetics; D008958:Models, Molecular; D011485:Protein Binding; D012709:Serum Albumin / Q000737:chemistry*; D013057:Spectrum Analysis; D013816:Thermodynamics; D014168:Transferrin / Q000737:chemistry*
chemical_list: D012709:Serum Albumin; D014168:Transferrin; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
keywords: FCCP; HSA; HTF; molecular modeling; spectroscopy