Protein–ligand
interaction studies are useful to determine
the molecular mechanism of the binding phenomenon, leading to the
establishment of the structure–function relationship. Here,
we report the binding of well-known antibiotic sulfonamide drugs (sulfamethazine,
SMZ; and sulfadiazine, SDZ) with heme protein myoglobin (Mb) using
spectroscopic, calorimetric, ζ potential, and computational
methods. Formation of a 1:1 complex between the ligand and Mb through
well-defined equilibrium was observed. The binding constants obtained
between Mb and SMZ/SDZ drugs were on the order of 104 M–1. SMZ with two additional methyl (−CH3) substitutions has higher affinity than SDZ. Upon drug binding,
a notable loss in the helicity (via circular dichroism) and perturbation
of the three-dimensional (3D) protein structure (via infrared and
synchronous fluorescence experiments) were observed. The binding also
indicated the dominance of non-polyelectrolytic forces between the
amino acid residues of the protein and the drugs. The ligand–protein
binding distance signified high probability of energy transfer between
them. Destabilization of the protein structure upon binding was evident
from differential scanning calorimetry results and ζ potential
analyses. Molecular docking presented the best probable binding sites
of the drugs inside protein pockets. Thus, the present study explores
the potential binding characteristics of two sulfonamide drugs (with
different substitutions) with myoglobin, correlating the structural
and energetic aspects.