One of the main differences in intercellular environment compared to laboratory condition is presence macromolecular crowders various compositions, sizes, and shapes. In this article, we have contemplated a systematic shape dependency on thermodynamics microsecond conformational fluctuation dynamics protein unfolding by taking human serum albumin (HSA) as model similar-sized crowders, namely, dextran-40, ficoll-70, PEG-35 different shapes, mimic cell environment. We observed that dextran-40...

A more compact structural conformation, higher active-site flexibility, lower viscosity, and solvent medium polarity are found to probably facilitate enzymatic activity in a hydrated deep eutectic (DES).

Over the past 20 years, most studied and debated aspect of macromolecular crowding is how it affects protein stability. Traditionally, explained by a delicate balance between stabilizing entropic effect or destabilizing enthalpic effect. However, this traditional theory cannot explain experimental observations like (i) negative (ii) entropy–enthalpy compensation. Herein, we provide evidence that associated water dynamics plays crucial role in controlling stability crowded milieu for first...

Macromolecular crowding bridges in vivo and vitro studies by simulating cellular complexities such as high viscosity limited space while maintaining the experimental feasibility. Over last two decades, impact of macromolecular on protein stability activity has been a significant topic study discussion, though still lacking thorough mechanistic understanding. This article investigates role associated water dynamics within crowded environments, using bromelain Ficoll-70 model systems....

The intercellular environment is known to be very different from the where most of elementary biological processes are studied in laboratory. As a result, there was considerable effort on cell mimicking either by confinement or introducing macromolecular crowding. In present study, dextran varying sizes has been used crowd protein, human serum albumin (HSA), and its structure, dynamics, activity were as function crowder concentration. By employing bulk single molecular level spectroscopic...

The cellular environment is crowded by macromolecules of various sizes, shapes, and charges, which modulate protein structure, function dynamics. Herein, we contemplated the effect three different macromolecular crowders: dextran-40, Ficoll-70 PEG-35 on active-site conformational dynamics, relative domain movement multi-domain human serum albumin (HSA). All crowders used in this study have zero charges similar sizes (at least dilute region) but shapes compositions. Some observations follow...

The mechanism of protein stabilization by osmolytes remains one the most important and long-standing puzzles. traditional explanation osmolyte-induced stability through preferential exclusion from surface has been seriously challenged observations like concentration-dependent reversal stabilization/destabilization. more modern stabilization/destabilization considers an indirect effect due to distortion water structure. It provides a general mechanism, but there are numerous examples...

A simple analytical model was constructed and validated to understand predict viscosity decoupling dynamic heterogeneity in solvent media. We assumed that the SE relationship is locally satisfied but their spatial average shows a breakdown.

Traditionally, deviation from Stokes–Einstein–Debye (SED) relation in terms of viscosity dependence medium dynamics, i.e., τx∝(ηT)p with p ≠ 1, is taken as a signature dynamic heterogeneity. However, it does not guarantee heterogeneity, the decoupling may also originate basic assumption SED. Here, we developed method to find stronger between (p 1) and heterogeneity rotational motion. Our approach exploited fact that heterogeneous media, solvatochromic probe will be solvated different extent...

Deep eutectic solvents (DESs) are new-generation media that can be fine-tuned to have desired properties circumventing economic and environmental issues. Typically, these ionic, only recently, nonionic DESs, having interesting properties, being explored. In this report, we examined the structure dynamics of a lauric acid/menthol (LA/Men) DES through steady-state emission, solvation dynamics, time-resolved fluorescence anisotropy, translational diffusion dynamics. The zero shift in emission...

Vulnerability to atmospheric conditions and their associated toxicity limit the practical/industrial use of perovskites despite tremendous promise in optoelectronics.

In this work, we have investigated the effects of denaturing agents, guanidine hydrochloride (GnHCl) and temperature, on overall structure, domain-I, domain-III human serum albumin (HSA) using circular dichroism (CD) spectroscopy steady-state, time-resolved fluorescence spectroscopy. We tagged Cys-34 HSA, located at N-(7-dimethylamino-4-methylcoumarin-3-yl)iodoacetamide Tyr-411 domain-III, p-nitrophenyl coumarin ester, for purpose. The CD studies reveal denaturation protein. follows expected...

Proteins are dynamic entity with various molecular motions at different timescale and length scale. Molecular crucial for the optimal function of an enzyme. It seems intuitive that these enzyme activity. However, it is not easy to directly correlate enzyme's dynamics activity due biosystems' enormous complexity. amongst many factors, structure two prime aspects combinedly control Therefore, having a direct correlation between protein straightforward. Herein, we observed correlated...

In this contribution, we have compared the stabilizing effect of sucrose on overall as well domain-II human serum albumin (HSA) against unfolding by different denaturating agents. HSA was denatured thermally raising temperature and also chemically guanidine hydrochloride (GnHCl) urea. Circular dichroism spectroscopy used to monitor change in structure HSA, whereas tryptophan fluorescence has been investigate local structural alteration within HSA. The degree folding using combinations...

Abstract Conformational heterogeneity is a defining characteristic of protein and vital in understanding its function folding landscape. In the present work, we interrogated presence conformational multi‐domain human serum albumin domain‐specific manner using red edge excitation shift (REES) native state also monitored variation along unfolding transition. We looked into origin such by varying solution viscosity. observed (1) even state, dynamics side chain exhibit varied behaviors depending...

Various biophysical techniques have been extensively employed to study protein aggregation due its significance. Traditionally, these methods detect at micrometer length scales and micromolar concentrations. However, unlike in vitro, typically occurs nanomolar concentrations vivo. Here, using fluorescence correlation spectroscopy (FCS), we captured bromelain as low ~20 nM, surpassing the detection limit of traditional like thioflavin T fluorescence, scattering, microscopy by more than one...

Hydrated deep eutectic solvents (DESs) are recognized for their potential in biocatalysis due to tunability, biocompatibility, greenness, and ability keep protein stable active. However, the mechanisms governing enzyme stability activity DES remain poorly understood. Herein, using bromelain as model acetamide (0.5)/urea(0.3)/sorbitol(0.2) DES, we provide experimental evidence that modulation of associated water plays a key role dictating hydrated DES. Specifically, rigid at higher...