Calorimetric observation of lysozyme degradation at elevated temperature in water and DMSO-water mixtures

Mukhametzyanov, Timur A.; Fatkhutdinova, Alisa A.; Sedov, Igor A.; Yakimova, Luidmila S.; Klimovitskii, Alexander E.

doi:10.1016/j.tca.2020.178826

Cited by 6 publications

(4 citation statements)

References 33 publications

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“…The faster scanning rates, available in FSC, not only allow to perform temperature-jump experiments to study the protein ensemble during folding, but have other important advantages, as well. The high heating rates limit the duration of the exposure of the protein solution to the high temperatures, thus decreasing possible contribution of the irreversible chemical degradation (e.g., deamidation [ 35 ]) or aggregation. Of course, if the protein solution is then rapidly cooled, the aggregation of the unfolded protein may occur (we have not detected it in the present work).…”

Section: Discussionmentioning

confidence: 99%

Refolding of Lysozyme in Glycerol as Studied by Fast Scanning Calorimetry

Fatkhutdinova

Mukhametzyanov

Schick

2022

IJMS

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The folding of lysozyme in glycerol was monitored by the fast scanning calorimetry technique. Application of a temperature–time profile with an isothermal segment for refolding allowed assessment of the state of the non-equilibrium protein ensemble and gave information on the kinetics of folding. We found that the non-equilibrium protein ensemble mainly contains a mixture of unfolded and folded protein forms and partially folded intermediates, and enthalpic barriers control the kinetics of the process. Lysozyme folding in glycerol follows the same or similar triangular mechanism described in the literature for folding in water. The unfolding enthalpy of the intermediate must be no lower than 70% of the folded form, while the activation barrier for the unfolding of the intermediate (ca. 140 kJ/mol) is about 100 kJ/mol lower than that of the folded form (ca. 240–260 kJ/mol).

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Section: Discussionmentioning

confidence: 99%

Refolding of Lysozyme in Glycerol as Studied by Fast Scanning Calorimetry

Fatkhutdinova

Mukhametzyanov

Schick

2022

IJMS

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“…and plants, serving as part of the innate immune response by protecting against bacteria and viruses. , Lysozyme can disrupt the bacterial wall due to its positive charge, and, in combination with antibiotics, can partially decrease the drug resistance due to a synergistic effect. , Furthermore, lysozyme is a monomeric protein, and its three-dimensional structure is stabilized by only four disulfide linkages. As a result, the denaturation process of lysozyme can be reversed relatively easily, − endowing lysozyme with relatively high stability against denaturation. Lysozyme has already been used in the delivery of hydrophobic drugs. − Due to the smaller size, lysozyme with conjugated drugs accumulates less likely in the kidney .…”

Section: Introductionmentioning

confidence: 99%

Mix and Shake: A Mild Way to Drug-Loaded Lysozyme Nanoparticles

Cao,

Tian,

et al. 2024

ACS Appl. Mater. Interfaces

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Biocompatible nanoparticles as drug carriers can improve the therapeutic efficiency of hydrophobic drugs. However, the synthesis of biocompatible and biodegradable polymeric nanoparticles can be time-consuming and often involves toxic solvents. Here, a simple method for protein-based stable drugloaded particles with a narrow polydispersity is introduced. In this process, lysozyme is mixed with hydrophobic drugs (curcumin, ellipticine, and dasatinib) and fructose to prepare lysozyme-based drug particles of around 150 nm in size. Fructose is mixed with the drug to generate nanoparticles that serve as templates for the lysozyme coating. The effect of lysozyme on the physicochemical properties of these nanoparticles is studied by transmission electron microscopy (TEM) and scattering techniques (e.g., dynamic light scattering (DLS) and small-angle X-ray scattering (SAXS)). We observed that lysozyme significantly stabilized the curcumin fructose particles for 7 days. Moreover, additional drugs, such as ellipticine and dasatinib, can be loaded to form dual-drug particles with narrow polydispersity and spherical morphology. The results also reveal that lysozyme dual ellipticine/dasatinib curcumin particles enhance the cytotoxicity and uptake on MCF-7 cells, RAW 264.7 cells, and U-87 MG cells due to the larger and rigid hydrophobic core. In summary, lysozyme in combination with fructose and curcumin can serve as a powerful combination to form protein-based stable particles for the delivery of hydrophobic drugs.

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“…A growing number of published studies report applications of fast thermal analyzes to micro- and nano-sized samples. These largely fall into two major categories—open chamber sensors for use with micro- or nano-sized solid materials or liquid droplets [ 32 ], e.g., for the analysis of proteins [ 33 , 34 ], or miniaturized enclosed micro-fluidic chambers [ 35 , 36 , 37 ], some within nanolitre range [ 38 ] or even as small as few picolitres [ 39 ].…”

Section: Introductionmentioning

confidence: 99%

“…Only a handful of dedicated in vivo studies have been published, mostly employing macroscopic preparations of bacterial cultures [ 46 ], with the notable exceptions of the direct real-time calorimetry study of an individual C. elegans worm [ 47 ] or unicellular Paramecium caudatum [ 48 ]. Some further advances were achieved with commercially available instruments such as Flash DSC [ 34 , 49 , 50 ] or TA Instruments [ 37 ] largely used to study lysozyme protein—an old favourite model protein continuously explored over the last 50 years. However, despite the consistent tendency towards miniaturization, simply scaling down traditional calorimetric techniques, although beneficial for material sciences, may not work well for biological objects, which cannot always be scaled down to suit instrument size or taken out of their environment.…”

Section: Introductionmentioning

confidence: 99%

AC/DC Thermal Nano-Analyzer Compatible with Bulk Liquid Measurements

et al. 2022

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Nanocalorimetry, or thermal nano-analysis, is a powerful tool for fast thermal processing and thermodynamic analysis of materials at the nanoscale. Despite multiple reports of successful applications in the material sciences to study phase transitions in metals and polymers, thermodynamic analysis of biological systems in their natural microenvironment has not been achieved yet. Simply scaling down traditional calorimetric techniques, although beneficial for material sciences, is not always appropriate for biological objects, which cannot be removed out of their native biological environment or be miniaturized to suit instrument limitations. Thermal analysis at micro- or nano-scale immersed in bulk liquid media has not yet been possible. Here, we report an AC/DC modulated thermal nano-analyzer capable of detecting nanogram quantities of material in bulk liquids. The detection principle used in our custom-build instrument utilizes localized heat waves, which under certain conditions confine the measurement area to the surface layer of the sample in the close vicinity of the sensing element. To illustrate the sensitivity and quantitative capabilities of the instrument we used model materials with detectable phase transitions. Here, we report ca. 106 improvement in the thermal analysis sensitivity over a traditional DSC instrument. Interestingly, fundamental thermal properties of the material can be determined independently from heat flow in DC (direct current) mode, by using the AC (alternating current) component of the modulated heat in AC/DC mode. The thermal high-frequency AC modulation mode might be especially useful for investigating thermal transitions on the surface of material, because of the ability to control the depth of penetration of AC-modulated heat and hence the depth of thermal sensing. The high-frequency AC mode might potentially expand the range of applications to the surface analysis of bulk materials or liquid-solid interfaces.

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Calorimetric observation of lysozyme degradation at elevated temperature in water and DMSO-water mixtures

Cited by 6 publications

References 33 publications

Refolding of Lysozyme in Glycerol as Studied by Fast Scanning Calorimetry

Refolding of Lysozyme in Glycerol as Studied by Fast Scanning Calorimetry

Mix and Shake: A Mild Way to Drug-Loaded Lysozyme Nanoparticles

AC/DC Thermal Nano-Analyzer Compatible with Bulk Liquid Measurements

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