Harmane, a naturally occurring beta-carboline alkaloid found in plants and animals, has sparked interest due to its potential biological impacts, specifically its neuro-pharmacological properties and its link with neurodegenerative ailments. This work studies the interaction between the bovine serum albumin (BSA) and the Harmane via multi-spectroscopic and computational analyses. Spectroscopic titrations at different temperatures and concentrations of the salt showed that Harmane forms a stable ground-state complex revealed by a static quenching of the fluorescence spectra. Thermodynamic characteristics of the binding show that BSA and Harmane have a 1:1 binding stoichiometry. The negative change in the Gibbs free energy (ΔG) indicates that the interaction is spontaneous, whereas the positive values of the change in entropy (ΔS) and the change in enthalpy (ΔH) indicate that hydrophobic forces predominate in the binding. The protein surface hydrophobicity index confirms significant surface hydrophobicity. The study also demonstrates the impact of metal ions on protein structure. According to molecular docking studies, Harmane binds to BSA in site II, domain III, subdomain IIIA. Molecular Dynamic (MD) modeling was used to reconfirm the perturbation in the stability and structure of the protein due to Harmane binding at different time frames.