title: Antidepressant Fluoxetine Modulates the In Vitro Inhibitory Activity of Buffalo Brain Cystatin: A Thermodynamic Study Using UV and Fluorescence Techniques.
abstract: Cystatins constitute a superfamily of homologous proteins. The major role of cystatins is to regulate the unwanted proteolysis and to protect the organism against endogenous proteases released from lysosomes, invading microorganisms and parasites that use cysteine proteases to enter the body. Imbalance in regulation of proteolytic activity may lead to a wide range of human diseases. An enormous progress has been made in understanding of protein degradation process under normal and pathological conditions; infact proteases are now clearly viewed as important drug targets. Fluoxetine a selective serotonin reuptake inhibitor (SSRI) is an antidepressant. It is used to treat major depressive disorders. In the present study binding of fluoxetine to cystatin was studied by UV and fluorescence quenching technique. Intrinsic fluorescence of fluoxetine complexed with purified buffalo brain cystatin (BC) was measured by selectively exciting the tryptophan residues. Gradual quenching was observed on complex formation. When cystatin was added to fluoxetine solutions at a molar ratio of 1 : 0.5, it not only quenched more than half of its fluorescence but also reduced the activity of cystatin. Stern-Volmer plots obtained from experiments carried out at 25(°)C showed the quenching of fluorescence to be a collisional phenomenon. Our results suggest the prime binding site for fluoxetine on BC to be at or near tryptophan residues. Fluoxetine quenched the fluorescence by a static process, which specifically indicates the formation of a complex.
authors.affiliation: Department of Biochemistry, Faculty of Life Sciences Aligarh Muslim University, Aligarh 202002, India.
authors.forename: Fakhra
authors.initials: F
authors.lastname: Amin
authors.affiliation: Department of Biochemistry, Faculty of Life Sciences Aligarh Muslim University, Aligarh 202002, India.
authors.forename: Bilqees
authors.initials: B
authors.lastname: Bano