Exploring the intermolecular interactions and contrasting binding of flufenamic acid with hemoglobin and lysozyme: A biophysical and docking insight

• Published in: International journal of biological macromolecules Vol. 116; pp. 1105 - 1118
• Main Authors: Ansari, Sameer Shakeel, Yousuf, Imtiyaz, Arjmand, Farukh, Siddiqi, Mohammad Khursheed, Naqvi, Saeeda
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.09.2018
• Subjects: Animals; Binding Sites; Cattle; Chickens; Flufenamic Acid - chemistry; Hemoglobins - chemistry; Molecular Docking Simulation; Muramidase - chemistry
• ISSN: 0141-8130; 1879-0003
• DOI: 10.1016/j.ijbiomac.2018.05.052

The intermolecular interaction of flufenamic acid (Hfluf) with two model proteins i.e., hemoglobin and lysozyme was explored using fluorescence, UV–vis, circular dichroism, DLS, and molecular docking techniques. The corroborative spectroscopic techniques suggested efficient binding of Hfluf to both the proteins. The S-V plot in Hb-Hfluf system showed positive deviation highlighting the presence of both static and dynamic quenching. Hence, ground state complex model and sphere of action quenching model were used for the study. In Lyz-Hfluf system, a linear S-V plot was obtained indicating the presence of a single quenching mechanism. FRET study suggested a high probability of energy transfer from Hb/Lyz to Hfluf. Our thermodynamic results revealed that binding reaction in both the systems was exothermic and spontaneous. The UV–vis spectroscopy demonstrated that the binding of Hfluf affected the globin, Soret and oxy-bands of Hb along with globin band and polypeptide backbone of Lyz. CD spectra revealed the enhancement of ɑ-helicity in Lyz and decrease in case of Hb whereas the Rh values of proteins from DLS experiment corroborated the CD findings. 3-D fluorescence spectra highlighted the conformational changes upon binding whereas docking studies predicted the active binding site of both the proteins as the binding site of Hfluf.