Etc 273t

Fluorescence experiments were performed by utilizing a JASCO FP 8300 spectrofluorometer (JASCO, Tokyo, Japan). Experiments were carried out at 25 °C in a 3 mm path-length quartz cuvette for 4 μM peptide in pH 7.0 buffer containing 100 mM KCl, 0.5 mM EDTA titrated with equimolar concentration of HTPu. The temperature of the cell holder was regulated by a JASCO ETC-273T temperature controller. Samples were prepared by same procedure. Excitation and emission slit width were 5 nm each and the samples were excited at 275 nm and the emission was recorded in a range of 300 nm to 500 nm. Experiment has been repeated in triplicates to reproduce the data. Modified Stern–Volmer equation was used to analyze fluorescence quenching data to find out various binding parameters for this interaction since binding parameters are vital to study about the binding mechanism.where the highest fluorescence intensity in the absence of ligand is F₀ whilst F depicts fluorescence intensity in the presence of ligand, K depicts the binding constant, n depicts the number of binding sites, and the concentration of RT is depicted by C.