Therapeutic Porphyrin “Hemin” Binds and Unfolds Hen Egg White Lysozyme: Experimental and In Silico Approaches to Study the Interaction

Abstract Hemin is an important iron‐containing porphyrin that has important therapeutic properties to control the symptoms of porphyria attacks that include pain, high blood pressure, rapid heartbeat, mental changes, and so forth. Lysozyme has many applications, which include from therapeutic to preservation of foods. The interaction of hemin with an important model protein, hen egg white lysozyme (HEWL), has been observed in this study through hybrid in silico with biophysical studies. There was a strong 1:1 cooperative binding between hemin and HEWL, which was revealed from the strong quenching of the latter by the former. The binding was favored by the decrease in temperature, which is due to the involvement of static type of quenching mechanism in the interaction. Experimental analyses suggested that the major forces involved in the binding were polar ones, and there was an efficient energy transfer from the fluorophore to the ligand. The combination of far‐UV CD spectroscopy and Rayleigh light scattering experiments established that hemin has the strong tendency to unfold HEWL. An extensive molecular docking investigation involving 2000 docking runs recognized the preferred binding site of hemin within HEWL, and also suggested that apart from the polar forces, hydrophobic interactions have also played an important role in the binding.