Refolding of Lysozyme in Glycerol as Studied by Fast Scanning Calorimetry

Fatkhutdinova, Alisa A.; Mukhametzyanov, Timur A.; Schick, Christoph

doi:10.3390/ijms23052773

Cited by 7 publications

(3 citation statements)

References 35 publications

(58 reference statements)

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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%

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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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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

et al. 2022

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“…Mukhametzyanov et al [ 6 ] study refolding of hen egg-white lysozyme in glycerol using a recently emerged method of fast scanning calorimetry. This method allows to work at millisecond timescales and provides a unique view of the non-equilibrium protein states.…”

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confidence: 99%

Recent Advances in Protein–Protein Interactions

Sedov

Zuev²

2023

IJMS

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Ultrasound-Assisted High-Voltage Cold Atmospheric Plasma Treatment on the Inactivation and Structure of Lysozyme: Effect of Treatment Voltage

Nasiru

Boateng

Wang

et al. 2022

Food Bioprocess Technol

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Emerging non-thermal processing techniques are gaining much attention due to their numerous advantages over traditional methods. Thus, in this study, we employed high voltage cold atmospheric plasma (HV-CAP) separately and synergetically with ultrasound on lysozyme to investigate its activity and structure. The HV-CAP was operated at 120, 140, and 160 kV for 3 min at a frequency of 120 Hz. A signi cant difference was observed between the untreated and both treatments in lysozyme activity, with a more pronounced result in the ultrasound-assisted HV-CAP treatment. Both treatments in uenced the tertiary structure of the lysozyme as lipid oxidation, carbonyl content, UV spectra, intrinsic uorescence, hydrophobicity, and sulfhydryl groups increased with increased voltage. Particle size distribution of lysozyme shows that noncovalent interactions affected the tertiary structure. The effects of both treatments on the secondary structure of lysozyme were demonstrated by FTIR-ATR, CD, and relative molar ellipticity. The results indicated that plasma-generated reactive species and ultrasound-induced cavitation in uenced the unfolding of tertiary and secondary structures of the lysozyme. The CD spectra postulated that lysozyme inactivation was closely linked to the α-helices and β-sheets components of the secondary structure. This study presents an original novel knowledge on the modi cation of activity and structure of lysozyme subjected to ultrasound-assisted HV-CAP treatment that has been missing from the literature.

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Refolding of Lysozyme in Glycerol as Studied by Fast Scanning Calorimetry

Cited by 7 publications

References 35 publications

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

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

Recent Advances in Protein–Protein Interactions

Ultrasound-Assisted High-Voltage Cold Atmospheric Plasma Treatment on the Inactivation and Structure of Lysozyme: Effect of Treatment Voltage

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