Thermal denaturation and protein stability analysis of Haliotis rubra hemocyanin

Marshall, Gavin; Valtchev, Peter; Dehghani, Fariba; Gomes, Vincent G.

doi:10.1007/s10973-015-4827-2

Cited by 10 publications

(9 citation statements)

References 22 publications

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“…The thermal denaturation of the Hc isoforms α D + N -HaH also was found to be irreversible ( Figure 1(b) , line C). Irreversibility of the thermal denaturation has been observed in DSC measurements of all studied to date Hcs [ 11 – 14 , 27 ] and limits the use of a standard equilibrium thermodynamic analysis.…”

Section: Resultsmentioning

confidence: 99%

“…The thermal denaturation of Hcs from different species has been described by the two-state irreversible model, and the parameters of the Arrhenius equation have been calculated, allowing comparison of transitions during Hc denaturation [ 11 – 13 , 26 , 27 ]. The activation energy value of 302.0 ± 6 (kJ·mol −1 ) and 596.0 ± 10 (kJ·mol −1 ) was determined for β -HaH and α D + N -HaH, respectively, which is in good agreement with values reported for Hcs in related species ( Table 3 ).…”

Section: Resultsmentioning

confidence: 99%

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Calorimetric Study of Helix aspersa Maxima Hemocyanin Isoforms

Todinova

Raynova

Idakieva

2018

Journal of Analytical Methods in Chemistry

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The thermal unfolding of hemocyanin isoforms, β-HaH and αD+N-HaH, isolated from the hemolymph of garden snails Helix aspersa maxima, was studied by means of differential scanning calorimetry (DSC). One transition, with an apparent transition temperature (Tm) at 79.88°C, was detected in the thermogram of β-HaH in 20 mM HEPES buffer, containing 0.1 M NaCl, 5 mM CaCl2, and 5 mM MgCl2, pH 7.0, at scan rate of 1.0°C min−1. By means of successive annealing procedure, two individual transitions were identified in the thermogram of αD+N-HaH. Denaturation of both hemocyanins was found to be an irreversible process. The scan-rate dependence of the calorimetric profiles indicated that the thermal unfolding of investigated hemocyanins was kinetically controlled. The thermal denaturation of the isoforms β-HaH and αD+N-HaH was described by the two-state irreversible model, and parameters of the Arrhenius equation were calculated.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Calorimetric Study of Helix aspersa Maxima Hemocyanin Isoforms

Todinova

Raynova

Idakieva

2018

Journal of Analytical Methods in Chemistry

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“…The parameters characterizing the process of irreversible thermal denaturation of HlH are respectively: an apparent T m 82.3 C; a calorimetric enthalpy (DH cal ) 108.7 MJ mol À1 . The irreversibility of the thermal denaturation was observed in the DSC measurements of all of the studied Hcs to date [13][14][15][16][17]31]. Kinetic models have been applied to analyze heat capacity curves if the model-derived transition temperatures and enthalpies are dependent on the heating rate [22,32,33].…”

Section: Irreversibility Of the Thermal Denaturationmentioning

confidence: 99%

“…1). Previous studies have applied this simplest model in the interpretation of the DSC data for the thermal denaturation of Hcs from different species [13][14][15]17,31]. To analyze the excess heat capacity functions, we fitted the data to the two-state irreversible model according to Equation (3) (Figures 4 and 6).…”

Section: Analysis Of the Calorimetric Datamentioning

confidence: 99%

Biophysical characterization of the structural stability of Helix lucorum hemocyanin

Idakieva

Todinova

Dolashki

et al. 2020

Biotechnology & Biotechnological Equipment

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The structural stability of the hemocyanin purified from the hemolymph of garden snails Helix lucorum (HlH) was investigated by means of far-UV circular dichroism (CD), differential scanning calorimetry (DSC) and transmission electron microscopy (TEM). For the first time, TEM analyses showed the presence of tubular polymers in hemocyanins after three-day dialysis against a stabilizing buffer containing high concentrations of Ca 2þ and Mg 2þ ions (100 mmol L À1 ). The conformational stability study of native HlH by means of CD in a wide pH range (2.5-11.5) defined the pH stability region of HlH at pH 6.5 À 8.0. DSC analyses demonstrated the thermal stability of this hemocyanin. One transition, with an apparent transition temperature (T m ) at 82.3 C, was detected in the heat capacity curve of HlH in 50 mmol L À1 Tris-HCl buffer, pH 7.2, at a heating rate of 1.0 C min À1 . The calorimetrically observed thermal transition correlates well with the unfolding transition monitored by CD measurements. The two-state kinetic model was used to analyse the process of irreversible thermal denaturation of HlH; ð a of 451 ± 4 kJ mol À1 was calculated. The obtained results on the conformational stability of HlH will facilitate the further investigation of the properties and potential biomedical applications of this respiratory protein.

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“…Additionally, due to its nutritious nature, mainly at lower dilutions, for the recombinant cell lines it has been used as an additive medium for the production of recombinant therapeutic protein in high-density perfusion culture of mammalian cells [5]. The stability of proteins at higher temperature is a major issue that hampers its applications in many targeted areas [6,7]. Bischof et al reported that it is imperative to understand that how protein loses stability and to what extent one can control this through the thermal environment as well as through chemical and mechanical modification of the protein structure [8].…”

Section: Introductionmentioning

confidence: 99%

Physicochemical Evaluation of Biofield Treated Peptone and Malmgren Modified Terrestrial Orchid Medium

Trivedi¹,

Branton²,

Trivedi³

et al. 2015

BIO

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Peptone and Malmgren modified terrestrial orchid (MMTO) has been used as a growth medium for tissue culture applications. This research study was conducted to explore the influence of Mr. Trivedi's biofield energy treatment on physicochemical properties of peptone and MMTO. The study was performed in two groups i.e. control and treated. The control group was kept aside as untreated, and the treated group was received the biofield energy treatment. The control and treated samples were further subjected to characterization by X-ray diffraction (XRD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), Fourier transform infrared (FT-IR) spectroscopy, particle size analyzer and surface area analyzer. The XRD analysis revealed the amorphous nature of the control and treated peptone samples. The DSC analysis showed an increase in thermal denaturation temperature of the treated peptone (196.22°C) as compared to the control sample (141.20°C). Additionally, the exothermic peak of treated sample (280°C) was increased as compared to the control (270°C). The DSC of control and treated MMTO showed the absence of the melting temperature in their respective DSC thermograms. The TGA analysis of the treated peptone showed an increase in onset of thermal degradation (172°C) with respect to the control (170°C). Nevertheless, the TGA thermogram of the treated MMTO (293.96°C) showed an increase in maximum thermal degradation temperature (T max ) as compared with the control (281.41°C). It indicated the good thermal stability of the treated peptone and MMTO samples. The FT-IR result of the treated peptone showed an upward shift in C-H (2817→2833 cm ) bands as compared to the control. Particle size analysis of the treated peptone showed an increase in d 50 (average particle size) and d 99 (size exhibited by 99% of particles) by 9.3 and 41.4%, respectively with respect to the control. Surface area analysis showed increase in surface area by 4.3% in the treated peptone. Altogether, the results corroborated that the biofield energy treatment had altered the physical, thermal and spectral properties of peptone and MMTO. It is assumed that biofield treated peptone and MMTO could be utilized as potential candidates for cell culture applications.

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Thermal denaturation and protein stability analysis of Haliotis rubra hemocyanin

Cited by 10 publications

References 22 publications

Calorimetric Study of Helix aspersa Maxima Hemocyanin Isoforms

Calorimetric Study of Helix aspersa Maxima Hemocyanin Isoforms

Biophysical characterization of the structural stability of Helix lucorum hemocyanin

Physicochemical Evaluation of Biofield Treated Peptone and Malmgren Modified Terrestrial Orchid Medium

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