A comprehensive insight into binding of hippuric acid to human serum albumin: a study to uncover its impaired elimination through hemodialysis

• Published in: PloS one 2013-08, Vol.8 (8), p.e71422
• Main Authors: Zaidi, Nida, Ajmal, Mohammad Rehan, Rabbani, Gulam, Ahmad, Ejaz, Khan, Rizwan Hasan
• Format: Article
• Language: English
• Subjects: Acids; Albumin; Binding; Binding Sites; Biochemistry; Biology; Biotechnology; Calorimetry; Calorimetry, Differential Scanning; Circular Dichroism; Dialysis; Dichroism; Differential scanning calorimetry; Enthalpy; Esterase; Fluorescence; Fluorescence spectroscopy; Heat measurement; Hemodialysis; Hippurates - chemistry; Hippuric acid; Human serum albumin; Humans; Hydrogen; Hydrogen Bonding; Hydrophobic and Hydrophilic Interactions; Hydrophobicity; Interdisciplinary aspects; Kinetics; Ligands; Molecular docking; Molecular Docking Simulation; Physics; Protein Binding; Proteins; Renal Dialysis; Serum albumin; Serum Albumin - chemistry; Sodium; Spectrometry, Fluorescence; Spectroscopy; Spectrum analysis; Static Electricity; Studies; Thermodynamics; Titration; Titration calorimetry; Toxins
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0071422

Binding of hippuric acid (HA), a uremic toxin, with human serum albumin (HSA) has been examined by isothermal titration calorimetry (ITC), differential scanning calorimetry (DSC), molecular docking, circular dichroism (CD) and fluorescence spectroscopy to understand the reason that govern its impaired elimination through hemodialysis. ITC results shows that the HA binds with HSA at high (K b ~10(4)) and low affinity (K b ~10(3)) sites whereas spectroscopic results predict binding at a single site (K b~10(3)). The HA form complex with HSA that involves electrostatic, hydrogen and hydrophobic binding forces as illustrated by calculated thermodynamic parameters. Molecular docking and displacement studies collectively revealed that HA bound to both site I and site II; however, relatively strongly to the later. Esterase-like activity of HSA confirms the involvement of Arg410 and Tyr411 of Sudlow site II in binding of HA. CD results show slight conformational changes occurs in the protein upon ligation that may be responsible for the discrepancy in van't Hoff and calorimetric enthalpy change. Furthermore, an increase in T(1)(m) and T(2)(m) is observed from DSC results that indicate increase in stability of HSA upon binding to HA. The combined results provide that HA binds to HSA and thus its elimination is hindered.