Exploring the fluorescence quenching interaction of amino acids and protein with natural organic matter by a multi-spectroscopic method

Abstract

The main objective of this research was to explore the fluorescence quenching mechanism of humic substance (Suwannee River natural organic matter, (SWNOM)) to amino acids (tryptophan, tyrosine) and protein (bovine serum albumin, (BSA)) by multi-spectroscopic methods. The locations of the peak of tryptophan, tyrosine, and BSA from the Parallel Factor Analysis were at Ex/Em 280/356 nm, 275/302 nm, and 280/344 nm, respectively. For SWNOM, two peaks appeared at Ex/Em of 240/448 nm, and 350/450 nm. Static quenching was the dominant quenching mechanism between BSA and SWNOM, whereas, no quenching was observed between tryptophan or tyrosine and SWNOM. Fourier-transform infrared spectroscopy and thermodynamic calculation demonstrated that hydrogen bonding and van der Waals force are the potential binding forces of BSA-SWNOM complex, as a result of rearrangement in the secondary polypeptide carbonyl hydrogen bonding network of BSA. This rearrangement led to the conformational change in BSA that induced quenching of BSA fluorescence by SWNOM.

Construing the Interactions of Coumarin Derivatives with the Main Protease of Stomach Pepsin Using Spectroscopic and Computational Analyses: Insights into the Binding Thermodynamics, Antifibrillation Studies, and Enzymatic Assay

“…However, based on the mechanism involved, quenching can be static, dynamic, or a combination of both. The specific quenching mechanism attributed to a particular interaction can be inferred by examining the temperature-dependent behavior, viscosity, and particularly the lifetime analysis of the excited state . The obtained spectra were corrected for the inner filter effect using eq , , as both 4-MU and UMB exhibit significant absorbance at the emission and excitation wavelengths of pepsin due to their fluorescence properties. F corr = F obs × 10 ( A em + A ex ) / 2 Here, the observed fluorescence intensity is represented by F obs , while the corrected fluorescence intensity is denoted as F corr .…”

“…The specific quenching mechanism attributed to a particular interaction can be inferred by examining the temperature-dependent behavior, viscosity, and particularly the lifetime analysis of the excited state. 44 The obtained spectra were corrected for the inner filter effect using eq 1a, 45,46 as both 4-MU and UMB exhibit significant absorbance at the emission and excitation wavelengths of pepsin due to their fluorescence properties.…”