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Increasing manufacturing and use of nanoparticles in industrial and biomedical applications creates the necessity to understand the impact of the interaction of nanoparticles with biomacromolecules. In the present study, graphene oxide nanosheets (GONS) were synthesized using modified Hummer's method and further characterized employing X-ray diffraction (XRD), UV, FTIR, and Raman spectroscopy. After characterization, the interaction of GONS with human serum albumin (HSA) was investigated to delineate the binding mechanism employing different kinds of spectroscopic techniques. Intrinsic fluorescence spectroscopy revealed that complex formation is taking place between HSA and GONS. Fluorescence-based binding studies suggested that GONS binds to HSA with a significant binding affinity, and the interaction is governed by dynamic quenching. The evaluation of enthalpy change (ΔH) and entropy change (ΔS) suggested that the HSA-GONS complex formation is driven by hydrogen bonding and van der Waals interaction and hence complexation process is seemingly specific. Structural transition in the microenvironment of HSA was monitored using synchronous fluorescence spectroscopy and three-dimensional fluorescence spectroscopy, which showed that GONS binding to HSA influences the microenvironment around tyrosine and tryptophan residues. Secondary structural alterations in HSA upon binding to GONS were measured using circular dichroism (CD) spectroscopy. Additionally, molecular docking provided an insight into the critical residues involved in HSA-GONS interaction and further validated our in vitro observations affirming interaction between GONS and HSA. The significance of this study is attributable to the fact that HSA and GONS can be used as nanocarriers in drug delivery systems., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Protein aggregation and glycation are directly associated with many pathological conditions including several neurodegenerative disorders. This study investigates the potential of naturally occurring plant product, Rosmarinic acid (RA), to inhibit the glycation and aggregation process. In this study, we report that varying concentrations of methylglyoxal (MG) induce advanced glycation end products (AGEs) and aggregates formation in HSA in vitro on day 6 and day 8, respectively. AGEs specific fluorescence confirmed the formation of AGEs in HSA in the presence of MG and further characterized the inhibitory potential of RA. It was found that the presence of RA prevented AGEs formation in vitro. Further, aggregates of HSA were characterized employing multi spectroscopic and microscopic techniques and RA was found to inhibit this process. This study proposes that RA could be a potential natural molecule to treat disorders where AGEs and aggregates of proteins play a pivotal role., Competing Interests: Declaration of competing interest None as per the best knowledge of authors., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Carbendazim is a benzimidazole fungicide used to control the fungal invasion. However, its exposure might lead to potential health problems. The present study evaluates the interaction of carbendazim (CAR) with human serum albumin (HSA) which is an important drug carrier protein and plays a very crucial role in the transportation of small molecules. A number of biophysical techniques were employed to investigate the binding of CAR with HSA. The increased UV-absorption of HSA on titrating with CAR suggests the formation of HSA-CAR complex and it could be due to the exposure of aromatic residues. The fluorescence study confirmed that CAR quenches the fluorescence of HSA and showed the static mode of quenching. CAR (50 µM) quenches around 56.14% of the HSA fluorescence. The quenching constant, binding constant, number of binding site and free energy change was calculated by fluorescence quenching experiment. Competitive displacement assay showed Sudlow's site I as the primary binding site of CAR on HSA. The synchronous fluorescence study revealed the perturbation in the microenvironment around tyrosine and tryptophan residues upon binding of CAR to HSA. The circular dichroism results suggested that the binding of CAR to HSA altered its secondary structure. Molecular docking experiment demonstrated the binding of CAR to Sudlow's site I of HSA. Docking studies suggested that the hydrogen bonding, van der Waals and pi-alkyl are playing role in the interaction of CAR with HSA. The study confirmed the conformational changes within HSA upon binding of CAR.

Serum protein glycation and formation of advanced glycation end products (AGEs) correlates with many diseases viz. diabetes signifying the importance of studying the glycation pattern of serum proteins. In our present study, methylglyoxal was investigated for its effect on the structure of human serum albumin (HSA); exploring the formation of AGEs and aggregates of HSA. The analytical tools employed includes intrinsic and extrinsic fluorescence, UV spectroscopy, far UV circular dichroism, Thioflavin T fluorescence, congo red binding, polyacrylamide gel electrophoresis (PAGE). UV and fluorescence spectroscopy revealed the structural transition of native HSA evident by new peaks and increased absorbance in UV spectra and quenched fluorescence in the presence of MG. Far UV CD spectroscopy revealed MG induced secondary structural alteration evident by reduced α-helical content. AGEs formation was confirmed by AGEs specific fluorescence. Increased ThT fluorescence and CR absorbance of 10mM MG incubated HSA suggests that glycated HSA results in the formation of aggregates of HSA. SEM and TEM were reported to have an insight of these aggregates. Molecular docking was also utilized to see site specific interaction of MG-HSA. This study is clinically significant as HSA is a clinically relevant protein which plays a crucial role in many diseases., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Amyloid fibrils are playing key role in the pathogenesis of various neurodegenerative diseases. Generally anionic molecules are known to induce amyloid fibril in several proteins. In this work, we have studied the effect of anionic food additive dye i.e., tartrazine (TZ) on the amyloid fibril formation of human serum albumins (HSA) and bovine serum albumin (BSA) at pHs7.4 and 3.5. We have employed various biophysical methods like, turbidity measurements, Rayleigh Light Scattering (RLS), Dynamic Light Scattering (DLS), intrinsic fluorescence, Congo red assay, far-UV CD, transmission electron microscopy (TEM) and atomic force microscopy (AFM) to decipher the mechanism of TZ-induce amyloid fibril formation in both the serum albumins at pHs7.4 and 3.5. The obtained results suggest that both the albumins forms amyloid-like aggregates in the presence of 1.0 to 15.0mM of TZ at pH3.5, but no amyloid fibril were seen at pH7.4. The possible cause of TZ-induced amyloid fibril formation is electrostatic and hydrophobic interaction because sulfate group of TZ may have interacted electrostatically with positively charged amino acids of the albumins at pH3.5 and increased protein-protein and protein-TZ interactions leading to amyloid fibril formation. The TEM, RLS and DLS results are suggesting that BSA forms bigger size amyloids compared to HSA, may be due to high surface hydrophobicity of BSA., (Copyright © 2017. Published by Elsevier B.V.)