Stability Of Myoglobin Research Articles

Spectroscopy-based analysis of the effect of Maillard reaction products on oxidative stability of carp myoglobin.

As a pro-oxidant, myoglobin (Mb) can induce lipid oxidation in meat. In this study, Fourier-transform infrared spectroscopy (FTIR), circular dichroism (CD), fluorescence spectroscopy, UV-vis spectrophotometry, and fluorescence microscopy were used to evaluate the impact of Maillard reaction products (MRPs) formed from glucose and lysine on the oxidative stability of carp Mb. MRPs were found to inhibit the auto-oxidation of Mb, reducing MetMb production by 8.45%. The static quenching of fluorescence indicated that MRPs interacted with Mb through hydrogen bonding and van der Waals forces, thereby enhancing the α-helical content by 11.57% and reducing the random coil content by 2.72%. The enhanced stability of this advanced structure helps to minimize the exposure of amino acid in the side chain and prevent the formation of MetMb. Fluorescence microscopy showed that MRPs reduce porphyrin ring degradation, consequently decreasing heme iron release. It is evident that MRPs play a crucial role in maintaining Mb structure stability and inhibiting its oxidation.

Impact of yohimbine on myoglobin stability: insights from molecular spectroscopic, and computational approaches

The prolific role of bioactive ligands in interacting with a variety of proteins has become a focal point of interest in pharmacokinetics and pharmacodynamics, thus sparking substantial enthusiasm within the realm of medicinal chemistry. The reversible binding of small molecules and proteins is a characteristic feature, and it’s essential to investigate these interactions to understand their mode and mechanism of action within the human body. Therefore, the primary objective of the present study is to understand the underlying mechanism by which yohimbine (Yoh) interacts with protein myoglobin (Mb), employing both in silico and in vitro methodologies. The emission spectroscopy studies yielded a binding constant of 105 M−1 and a binding site ratio of 1:1. The structural perturbation induced in the protein Mb by Yoh was also illustrated by circular dichroism. The results of the molecular docking investigation resulted in numerous significant interactions between Mb and Yoh, indicating a substantial binding affinity. The accuracy of the docking data was further confirmed through the use of molecular dynamics (MD) simulations, which were then followed by principal component analysis and free energy landscape investigations. The study posited that the stability of the Mb-Yoh complex remains intact throughout the simulated duration, exhibiting little alterations in its structural conformation. Therefore, the association between ligand–protein plays a key role in determining circulatory lifetimes and bioavailability. These factors, in turn, are pivotal in the rational drug design process.

Maillard reaction products inhibit lipid oxidation by regulating myoglobin stability in washed muscle model.

Lipid oxidation significantly contributes to muscle deterioration, with heme protein oxidation playing a crucial role in this process. This study investigated the inhibition of heme protein oxidation by Maillard reaction products (MRPs) using a washed muscle model combining washed carp with myoglobin (Mb). Protein oxidation products, protein texture, and lipid oxidation levels were assessed. Results showed that metmyoglobin (MetMb) is a primary driver of lipid oxidation in meat, likely due to Mb oxidation, exposing hydrophobic groups that bind to lipids. MRPs at concentrations of 0.5% and 1% effectively inhibited Mb oxidation. Specifically, treatment with 1% MRPs reduced MetMb formation by 15.38%, protein carbonyl content by 6.53%, hydroxyl radical content by 20.37%, protein aggregation by 40.81%, and particle size by 36.95% during later storage stages, thereby preserving Mb stability. In the Mb-mediated oxidation model, 1% MRPs inhibited the formation of primary and secondary oxidative metabolites by 59.47% and 68.19%, respectively, while maintaining muscle tissue texture integrity. PRACTICAL APPLICATION: The antioxidation of Maillard reaction products (MRPs) improves the stability of common carp. By inhibiting the autoxidation of myoglobin and lipid, MRPs help preserve the texture and color of fish muscle while extending its shelf life. This study provides a valuable reference for effectively controlling lipid oxidation in refrigerated fish products and enhancing their overall quality.

Beef production, physicochemical quality, oxidative shelf-life, fatty acid profile and sensory effects of replacing sorghum for maize in finisher diets

Climate change and high demand for maize have prompted the search for climate-smart, energy feedstuffs such as sorghum for use in beef finisher diets. Relative to maize, sorghum has comparable metabolizable energy content and contains more polyunsaturated fatty acids and bioactive phenolic compounds that may enhance beef health value, oxidative shelf-life, and sensory quality. The study investigated the impact of graded sorghum levels replacing maize in beef finisher diets on nutrient utilization, production, physicochemical quality, fatty acid composition, oxidative shelf-life, and sensory quality attributes of beef. Thirty-five, seven-months-old (230 ± 28 kg average initial weight) Angus steers were individually housed in pens and randomly allocated to five pellet diets formulated by replacing white maize in the basal diet (control) with 100, 200, 300, and 400 g/kg DM of sorghum. Diet did not affect (P > 0.05) nutrient intake, digestibility and utilization, growth performance, carcass characteristics, longissimus thoracis (LT) meat physicochemical quality, fatty acid contents and colour coordinates. Metmyoglobin and colour coordinates increased (P ≤ 0.05) with retail display except redness which declined (P ≤ 0.05). Lipid oxidation of LT meat aged for day 1 tended to increase (P ≤ 0.10) with increased dietary substitution of sorghum for maize in beef finishing diets. In addition, antioxidant activity, metallic aroma and liver-like flavour linearly increased (P ≤ 0.05) with sorghum inclusion in the diet. In conclusion, sorghum can fully replace maize in beef finisher diets with neutral effects on beef production, physicochemical quality, health value and colour stability, and somewhat desirable impact on antioxidant activity and myoglobin stability and undesirable effects on aroma and flavour of LT meat.

Colour stability improving of yellowfin tuna slices by compound antioxidant against oxidation of myoglobin

SummaryBrown stain is a major problem for tuna during storage. This study aimed to investigate the impact of antioxidants on myoglobin oxidation and the promotion of metmyoglobin (MetMb) reduction in tuna slices. Response surface methodology (RSM) was used to optimise the formula of the compound antioxidant in yellowfin tuna (YFT) slices, with the percentage of MetMb as the response value. The ratios of ε‐polylysine, sodium lactate and D‐sodium ascorbate were 0.43%, 2.21% and 2.71%, respectively. The interactions between sodium lactate and D‐sodium ascorbate had a greater impact on the percentage of MetMb. Then, verification experiments were conducted, and the results revealed that the samples treated with compound antioxidants had higher a* values, total sulfhydryl content, Ca2+‐ATPase activity, total reducing activity (TRA) and MetMb‐reducing activity (MRA) and lower percentages of MetMb and carbonyl contents than did the control samples. It also helps to maintain the stability of the NADH/NAD+ ratio, which contributes to facilitating MetMb reduction. The results showed that the compound antioxidant significantly inhibited myoglobin oxidation, promoted the reduction of MetMb in tuna slices and significantly enhanced the myoglobin stability of refrigerated YFT slices.

Effect mechanism of capsaicin and dihydrocapsaicin in chili on the oxidative stability of myoglobin in duck meat.

Myoglobin (Mb) in duck meat is commonly over-oxidized when heated at high temperatures, which may worsen the color of the meat. Enhancing the oxidative stability of Mb is essential for improving the color of duck meat. Capsaicin and dihydrocapsaicin (CA-DI) in chili exhibit antioxidant properties. This study investigated the effects of CA-DI on the structure and oxidative damage of Mb by fluorescence spectroscopy, differential scanning calorimetry analysis and particle size in duck meat during heat treatment. When the ratio of CA-DI to Mb was 10:1 g kg-1 and heat-treated for 36 min, oxymyoglobin significantly increased, and metmyoglobin significantly decreased compared with the control group (P < 0.05). In parallel, the carbonyl content of Mb in the CA-DI group decreased by 43.40 ± 0.10%, the sulfhydryl content increased by 188 ± 0.21%, and the free radical scavenging activity of Mb was significantly enhanced (P < 0.05). Moreover, the addition of CA-DI resulted in a significant decrease in the particle size of the Mb surface (P < 0.05). When the ratio of CA-DI to Mb was 10:1 g kg-1, CA-DI enhanced the thermal stability and significantly increased the thermal denaturation temperature of Mb. The molecular docking results indicated that hydrophobic interactions and hydrogen bonds were involved in the binding of CA-DI to Mb. CA-DI could combine with Mb and improve the oxidation stability of Mb in duck meat. This suggested that CA-DI could be a potential natural antioxidant that improves the color of meat products. © 2024 Society of Chemical Industry.

Study on the interaction mechanism between (−)-epigallocatechin-3-gallate and myoglobin: Multi-spectroscopies and molecular simulation

(−)-Epigallocatechin-3-gallate (EGCG) is remarkably efficacious in inhibiting the browning of red meat. We therefore propose a hypothesis that EGCG forms complexes with myoglobin, thereby stabilizing its structure and thus preventing browning. This study investigated the interaction mechanism between EGCG and myoglobin. EGCG induced static quenching of myoglobin. Noncovalent forces, including hydrogen bonds and van der Waals, primarily governing the interactions between myoglobin and EGCG. The interactions primarily disrupted myoglobin's secondary structure, thus significantly reducing surface hydrophobicity by 53% (P < 0.05). The modification augmented the solubility and thermal stability of myoglobin. The radius of gyration (Rg) value fluctuated between 1.47 and 1.54 nm, and the hydroxyl groups in EGCG formed an average of 2.93 hydrogen bonds with myoglobin. Our findings elucidated the formation of stable myoglobin–EGCG complexes and the myoglobin–EGCG interaction, thus confirming our initial hypothesis.

Molecular insights into the interaction between myoglobin and Imidacloprid: Multi-spectral experiments and computational simulations

Imidacloprid (IMI) belongs to the neonicotinoid insecticides, specifically the imidazole chemical group, which exhibits a high level of contact, digestive, and systemic effects on the central nervous system of insects. In this study, we investigated the impact of IMI on the conformation and structural stability of myoglobin (Mb) protein using spectroscopic techniques combined with molecular docking and simulation approaches. Additionally, we examined the thermal stability of the resulting complex. The UV–Visible results demonstrated the interaction between IMI and Mb protein. The intrinsic fluorescence of Mb was effectively quenched by IMI, indicating successful binding and the formation of a stable complex. Thermodynamic analysis revealed that the spontaneous bonding process was primarily governed by van der Waals forces and hydrogen bonds. FT-IR analysis indicated that IMI induced conformational changes in Mb, providing further evidence of its potential toxicity and structural damage to Mb. Furthermore, the interaction significantly enhanced the thermal stability of Mb compared to Mb alone. Molecular docking revealed the preferred binding site of IMI within the Mb structure. Molecular dynamics (MD) simulation results further supported the findings by demonstrating a decrease in compactness and an increase in the stability of the Mb-IMI complex. These findings offer valuable insights into the potential toxicity risks of IMI to human health and shed light on the binding mechanism between IMI and Mb. The results of this study serve as a useful model for gaining a deeper understanding of the effects of these pesticides on proteins and body health.

Investigation of the stability of myoglobin in the presence of amino acid ionic liquids

Investigation of the stability of myoglobin in the presence of amino acid ionic liquids

Stabilization of myoglobin from different species (produced by cellular agriculture) using food-grade natural and synthetic antioxidants

Cellular agriculture products, like myoglobin, are increasingly used by the food industry to provide desirable sensory properties to plant-based meat substitutes. This study elucidated the physicochemical properties and redox stability of myoglobin from both natural (equine) and cellular agriculture (bovine, sperm whale, and leopard) sources. The electrical characteristics and water-solubility of the different myoglobin samples were measured from pH 2.5 to 8.5. The isoelectric point of the myoglobin samples depended on the species, being pH 5.5 for equine, pH 4.5 for leopard and bovine, and pH 6.5 for sperm whale. All myoglobin samples had a solubility greater than 80% across the entire pH range studied. All myoglobin solutions appeared red and had two peaks in their UV–visible absorbance spectra after one day, which is consistent with oxymyoglobin formation. Equine myoglobin at pH 8 was selected to study its redox and color stability over time, where the oxymyoglobin oxidative status closely paralleled with the redness of the solutions. The effects of antioxidants (ascorbic acid, caffeic acid, catechin, gallic acid, quercetin, taxifolin, Trolox, and 4-methylcatechol) on the redox and color stability (redness) of the equine myoglobin (pH 8.0) was also studied. Antioxidants with low reduction potential values (ascorbic acid and quercetin) were particularly effective at enhancing the color stability of oxymyoglobin. The computational modeling study showed that amino acids on the myoglobin interacted with antioxidants through hydrogen bonds. The insights obtained may have important implications for the use of cellular agriculture to produce myoglobin for food applications.

Effect of magnetic field modification on oxidative stability of myoglobin in sarcoplasm systems

This study aimed to investigate the effect of magnetic fields (0, 3, 6, 12 mT) on the oxidation characteristics of myoglobin (Mb) in the sarcoplasmic protein (SP) system and to understander the underlying mechanism. The metmyoglobin content, Soret band of heme iron porphyrin, protein conformation and molecular weight distribution were measured in different Mb and SP samples. The results showed that the primary oxidation site of hydroxyl radical on Mb was likely to be the porphyrin ring structure and the side chain group of protein rather than the central iron atoms, what’s more, 12 mT magnetic field treatment had an inhibitory effect on the oxidative damage induced by hydroxyl radical.

Impact of Washing with Antioxidant-Infused Soda-Saline Solution on Gel Functionality of Mackerel (Auxis thazard) Surimi.

Mackerel (Auxis thazard), a tropical dark-fleshed fish, has the potential to be used in the production of surimi. It is necessary to identify the optimal washing method to make better use of this species since efficient washing is the most important step in surimi processing to ensure maximal gelling and high-quality surimi. The purpose of this study was to evaluate the combined effect of cold carbonated water (CW) with NaCl and antioxidants in washing media, so-called antioxidant-infused soda-saline solution, on lipid and myoglobin removal efficacy, biochemical characteristics, gelling properties, sensory features, and the oxidative stability of mackerel surimi in comparison with unwashed mince (T1) and conventional water washed surimi (T2). Mackerel mince was washed with CW in the presence of 0.6% NaCl at a medium to mince ratio of 3:1 (v/w) without antioxidant (T3) or with the addition of 1.5 mM EDTA plus 0.2% (w/v) sodium erythorbate and 0.2% sodium tripolyphosphate (T4), 100 mg/L gallic acid (T5), and 5 mM citric acid containing 8 mM calcium chloride (T6). During the first washing cycle, the antioxidants were mixed into the washing medium. The second and third washing cycles were then completed with cold water. The yields of all treatments were roughly 75-83%, based on the gross weight of the raw mince. The pH of the surimi was in a range of 5.47-6.46. All of the surimi had higher reactive sulfhydryl (SH) content and surface hydrophobicity but lower Ca2+-ATPase activity than unwashed mince (p < 0.05). After washing, lipids decreased significantly (p < 0.05), accounted for a 65-76% reduction. The T2 surimi had the highest peroxide value (PV). T1 had the lowest conjugated diene value. T1 and T4 surimi had the lowest TBARS value (p < 0.05). A lower non-heme iron level was found in all antioxidant-treated samples than in T1. Washing can increase the redox stability of myoglobin regardless of the washing media, as seen by the relatively low metmyoglobin levels. According to the dynamic viscoelastic behavior, all surimi and unwashed mince underwent the same degree of sol-gel transition following heat gelation. T1 showed the lowest breaking force, deformation, gel strength, and whiteness (p < 0.05). Surimi made from T4 or T5 had the highest gel strength when both breaking and deformation were considered, but the latter's expressible drip was noticeably higher. Surimi gel appears to be stabilized against lipid oxidation, as demonstrated by low PV and TBARS levels, when produced with T4. Because of the low level of TBARS, all 10 panelists rated rancid odor as low (~1 out of 4), with no significant variations across treatments. Only treatments with T4 and T6 tended to have a lower fishy odor score as compared to unwashed mince. Scanning electron microscope demonstrated that surimi gels washed with all washing media exhibited microstructures that were very comparable, with the exception of the T6 treatment, which had big pores and aggregates. Based on the quality features, T4 appeared to be the optimal medium to enhance the gel functionality of mackerel surimi.

Role of Stingray (Himantura signifier) Non-Protein Nitrogenous Fraction on the Oxidative Stability of Lipid and Myoglobin.

Non-protein nitrogen (NPN) is abundant in stingray (Himantura signifier) muscle, which also has in vitro antioxidant activity. In this study, NPN from stingray muscle was further investigated for its antioxidant properties in lecithin liposome and oxymyoglobin model systems to validate its protective impact against lipid and myoglobin oxidations during storage for 120 min at various temperatures (4, 25, and 60 °C). NPN solution (10 ppm nitrogen) was added to the lecithin liposome system at different concentrations (0, 0.5, 1, 5, and 10% (v/v)) to investigate its effects on lipid stability by measuring the conjugated diene (CD), peroxide value (PV), and thiobarbituric acid reactive substances (TBARS) contents. In the oxymyoglobin system, NPN solution (10 ppm nitrogen) was also added at different concentrations (0, 0.5, 1, 5, and 10% (v/v)) to the oxymyoglobin solution in order to examine its effect on the stability of myoglobin by determining the contents of oxymyoglobin, metmyoglobin, and protein carbonyl. According to the findings, in all NPN concentrations, the system incubated at 4 °C had the lowest levels of lipid oxidation as measured by CD, PV, and TBARS values, and the lowest levels of myoglobin oxidation. At all incubating temperatures, the oxymyoglobin and lipid oxidation of all model systems tended to rise with the lengthening of the incubation duration. With the addition of 5% NPN, however, the lowest CD, PV, TBARS, oxymyoglobin oxidation, metmyoglobin formation, and protein carbonyl content were all observable, and the remarkable result was discovered during incubation at 4 °C. The results indicate that stingray NPN, especially at 5%, can be used to delay lipid and myoglobin oxidation, particularly at 4 °C. In order to prolong the shelf life of products with dark-fleshed fish and red meat, stingray NPN might be used as an alternative antioxidant to delay the oxidation of lipid and myoglobin during cold chain storage.

Structural insights into the binding behavior of NiO with myoglobin

Valuable and useful information on the biotechnological and medical applications of nanoparticles can be achieved through the interactions of proteins with nanoparticles and the structural changes exerted by nanoparticles on these biomolecules. Nickel oxide (NiO) nanoparticles have widespread industrial and therapeutic applications in the medical and biological sciences. Therefore, structural changes and stability exerted by NiO nanoparticles on myoglobin as a globular protein sample were examined at a pH of 7.4 using UV–Vis spectroscopy and fluorescence emission techniques. Myoglobin is a monomeric protein that functions as oxygen storage in the skeletal muscle of vertebrates. Theoretical molecular docking technique was also applied to predict the interactions between myoglobin and NiO nanoparticle complex. Results showed that binding of NiO nanoparticles to myoglobin quenched its emission signals through a static quenching mechanism. Also, electrostatic interaction was predominating in the complex formation relative to other non-covalent interactions. Besides, the spontaneous bonding of NiO nanoparticles was confirmed by negative Gibbs free energy (ΔG°). Measurements performed by protein thermal stability assay showed that thermal stability of myoglobin increased with rising concentrations of NiO nanoparticles. CD spectroscopic studies showed that in the presence of NiO nanoparticles, the content of α-helix protein increased and the content of other secondary structures decreased. Accordingly, it is clearly evident that the structure and stability of myoglobin are altered based on the experimental and theoretical data obtained under the influence of NiO nanoparticles.

Structural Refolding and Thermal Stability of Myoglobin in the Presence of Mixture of Crowders: Importance of Various Interactions for Protein Stabilization in Crowded Conditions.

The intracellular environment is overcrowded with a range of molecules (small and large), all of which influence protein conformation. As a result, understanding how proteins fold and stay functional in such crowded conditions is essential. Several in vitro experiments have looked into the effects of macromolecular crowding on different proteins. However, there are hardly any reports regarding small molecular crowders used alone and in mixtures to observe their effects on the structure and stability of the proteins, which mimics of the cellular conditions. Here we investigate the effect of different mixtures of crowders, ethylene glycol (EG) and its polymer polyethylene glycol (PEG 400 Da) on the structural and thermal stability of myoglobin (Mb). Our results show that monomer (EG) has no significant effect on the structure of Mb, while the polymer disrupts its structure and decreases its stability. Conversely, the additive effect of crowders showed structural refolding of the protein to some extent. Moreover, the calorimetric binding studies of the protein showed very weak interactions with the mixture of crowders. Usually, we can assume that soft interactions induce structural perturbations while exclusion volume effects stabilize the protein structure; therefore, we hypothesize that under in vivo crowded conditions, both phenomena occur and maintain the stability and function of proteins.

Hofmeister effects on protein stability are dependent on the nature of the unfolded state.

The interpretation of a salt's effect on protein stability traditionally discriminates low concentration regimes (<0.3 M), dominated by electrostatic forces, and high concentration regimes, generally described by ion-specific Hofmeister effects. However, increased theoretical and experimental studies have highlighted observations of the Hofmeister phenomena at concentration ranges as low as 0.001 M. Reasonable quantitative predictions of such observations have been successfully achieved throughout the inclusion of ion dispersion forces in classical electrostatic theories. This molecular description is also on the basis of quantitative estimates obtained resorting to surface/bulk solvent partition models developed for ion-specific Hofmeister effects. However, the latter are limited by the availability of reliable structures representative of the unfolded state. Here, we use myoglobin as a model to explore how ion-dependency on the nature of the unfolded state affects protein stability, combining spectroscopic techniques with molecular dynamic simulations. To this end, the thermal and chemical stability of myoglobin was assessed in the presence of three different salts (NaCl, (NH4)2SO4 and Na2SO4), at physiologically relevant concentrations (0-0.3 M). We observed mild destabilization of the native state induced by each ion, attributed to unfavorable neutralization and hydrogen-bonding with the protein side-chains. Both effects, combined with binding of Na+, Cl- and SO42- to the thermally unfolded state, resulted in an overall destabilization of the protein. Contrastingly, ion binding was hindered in the chemically unfolded conformation, due to occupation of the binding sites by urea molecules. Such mechanistic action led to a lower degree of destabilization, promoting surface tension effects that stabilized myoglobin according to the Hofmeister series. Therefore, we demonstrate that Hofmeister effects on protein stability are modulated by the heterogeneous physico-chemical nature of the unfolded state. Altogether, our findings evidence the need to characterize the structure of the unfolded state when attempting to dissect the molecular mechanisms underlying the effects of salts on protein stability.

Organic solvent stability and long-term storage of myoglobin-based carbene transfer biocatalysts.

Recent years have witnessed a rapid increase in the application of enzymes for chemical synthesis and manufacturing, including the industrial-scale synthesis of pharmaceuticals using multienzyme processes. From an operational standpoint, these bioprocesses often require robust biocatalysts capable of tolerating high concentrations of organic solvents and possessing long shelflife stability. In this work, we investigated the activity and stability of myoglobin (Mb)-based carbene transfer biocatalysts in the presence of organic solvents and after lyophilization. Our studies demonstrate that Mb-based cyclopropanases possess remarkable organic solvent stability, maintaining high levels of activity and stereoselectivity in the presence of up to 30%-50% (v/v) concentrations of various organic solvents, including ethanol, methanol, N,N-dimethylformamide, acetonitrile, and dimethyl sulfoxide. Furthermore, they tolerate long-term storage in lyophilized form, both as purified protein and as whole cells, without significant loss in activity and stereoselectivity. These stability properties are shared by Mb-based carbene transferases optimized for other type of asymmetric carbene transfer reactions. Finally, we report on simple protocols for catalyst recycling as whole-cell system and for obviating the need for strictly anaerobic conditions to perform these transformations. These findings demonstrate the robustness of Mb-based carbene transferases under operationally relevant conditions and should help guide the application of these biocatalysts for synthetic applications.

Effects of Ethylene Glycol on the Structure and Stability of Myoglobin Using Spectroscopic, Interaction, and In Silico Approaches: Monomer Is Different from Those of Its Polymers.

Investigation of changes in thermal stabilities and structures of proteins in the presence of different co-solutes (ligands) is an integral part in the basic research, discovery, and development of drugs. Ethylene glycol (EG) is known to be toxic and causes teratogenic, inducing primarily skeletal and external malformations and other diseases. The effect of EG on the structure and thermal stability of myoglobin (Mb) was studied using various spectroscopic techniques at pH 7.0 and two different temperatures. As revealed by circular dichroism, Trp fluorescence, nano-DSF, and absorption (UV and visible) measurements, EG (i) has no significant effect on secondary and tertiary structures of Mb at 25 °C, and (ii) it decreases the thermal stability of the protein, which increases with increasing concentration of EG. As revealed by ANS (8-anilino-1-naphthalene sulfonic acid) fluorescence measurements, heat-induced denatured protein has newly exposed hydrophobic patches that bind to ANS. Isothermal titration calorimetry revealed that the interaction between EG and Mb is temperature dependent; the preferential interaction of EG is entropy driven at low temperature, 298 K (25 °C), and it is enthalpy driven at higher temperature, 343 K (70 °C). Molecular docking study showed that EG interacts with side chains of amino acid residues of Mb through van der Waals interactions and hydrogen bonding.

Targeted Myoglobin Delivery as a Strategy for Enhancing the Sensitivity of Hypoxic Cancer Cells to Radiation.

SummaryThe effectiveness of cancer radiotherapy is frequently hindered by the hypoxia of the tumor microenvironment. Direct delivery of oxygen to hypoxic tumor tissues is an attractive strategy to overcome this hypoxia-associated radioresistance. Herein, we report the generation of submicron-sized particles comprising myoglobin fused to the crystal-forming domain of Cry3Aa protein for the targeted delivery of oxygen to cancer cells. We demonstrate that myoglobin-containing particles were successfully produced in Bacillus thuringiensis with the assistance of the Cry3Aa domain I. Furthermore, these particles could be genetically modified to incorporate the cell penetrating peptide TAT and cell targeting peptide A549.1, resulting in particles that exhibited improved cellular uptake and targeting toward A549 cells. Notably, these myoglobin-containing particles increased the intracellular oxygen levels of A549 cells and thereby sensitized them to radiation. These findings suggest that the targeted delivery of O2-bound myoglobin could be an effective approach to enhance the efficacy of radiotherapy.

Insight into the binding of glycerol with myoglobin: Spectroscopic and MD simulation approach

Stability of proteins plays a significant role not only in their biological function but also in medical science and protein engineering. Since proteins are only stable in special conditions, maintaining their stability and function in biological and biotechnological applications may pose serious challenges. Osmolytes provide a general method of shielding proteins from the unfolding and aggregation caused by extreme stress on the environment. In such studies, the researchers used spectroscopic and simulation approaches to study the alterations of the myoglobin structure and stability in glycerol presence. Experimental results showed a stability improvement of the complex myoglobin-glycerol. After the addition of glycerol resulting in the initiation of hydrogen bonds and higher levels of hydrophobicity, the increase of the Tm was observed. The static mode quenching observed in this study. Van der Waals forces and hydrogen bindings had a decisive and significant role concerning the stability of protein which was consistent with the modeling results. Molecular dynamics simulation showed that the glycerol presence could enhance myoglobin stability. The consistency between the theoretical studies and experimental findings demonstrates that the method proposed in this study could provide a useful method for protein-ligand complex investigations.