Study of microheterogeneous environment of protein Human Serum Albumin by an extrinsic fluorescent reporter: a spectroscopic study in combination with Molecular Docking and Molecular Dynamics Simulation

Journal of Photochemistry and Photobiology. B, Biology
Sankar JanaNikhil Guchhait

Abstract

We report extrinsic fluorescent probe 5-(4-dimethylamino-phenyl)-penta-2,4-dienoic acid (DMAPPDA) as a molecular reporter for studying microheterogeneous environment of protein Human Serum Albumin (HSA) via spectral modification of the probe under physiological condition. Steady state emission, fluorescence anisotropy, Red Edge Excitation Shift (REES), far-UV Circular Dichroism (CD), Atomic Force Microscopy (AFM) imaging, time resolved spectral measurements, Molecular Docking and Molecular Dynamics (MD) Simulation techniques have been used to fulfill this achievement. Interaction of the probe with HSA is signaled by the blue shift of the fluorophore emission maxima with enhancement of fluorescence intensity. The increase in steady state anisotropy, REES and fluorescence lifetime values with increasing protein concentrations indicates interaction and movement of the probe from free aqueous media to the more restricted less polar hydrophobic interior of protein. Experimental results obtained from Benesi-Hildebrand plot support the formation of 1:1 HSA-DMAPPDA complex with high binding constant and negative free energy change. Thermal denaturation of the probe bound protein has also been tracked using the spectral response of DMAP...Continue Reading

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Citations

Oct 17, 2015·Journal of Biomolecular Structure & Dynamics·Jitendra Kumar MauryaRajan Patel
Aug 8, 2013·Journal of Biochemical and Molecular Toxicology·Junlong WuWeihua Ren
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Jan 15, 2014·Langmuir : the ACS Journal of Surfaces and Colloids·Uttam Anand, Saptarshi Mukherjee
Oct 6, 2021·Luminescence : the Journal of Biological and Chemical Luminescence·Yulin ZhuShujun Shang

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