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Food Chemistry May 2024The effects of oat β-glucan (OG) combined with ultrasound-assisted treatment on thermal aggregation behavior of silver carp myofibrillar protein (MP) under low salt...
The effects of oat β-glucan (OG) combined with ultrasound-assisted treatment on thermal aggregation behavior of silver carp myofibrillar protein (MP) under low salt concentration were investigated. The particle size and turbidity of MP were increased to higher levels by OG participation or ultrasound treatment during the two-stage heating. Both OG and ultrasonic treatment promoted the unfolding of MP structure, evidenced by the gradual decrease of α-helix content and fluorescence intensity, as well as the increase of β-sheet content, surface hydrophobicity and sulfhydryl content. Compared to solely OG or ultrasonic treatment, the combination of OG and ultrasound further promoted the unfolding of MP and more sulfhydryl groups were exposed in the pre-heating stage, which was conducive to strengthen the chemical forces between MP molecules. Additionally, AFM analysis revealed that the apparent morphology of the OG combined with ultrasonic treated group exhibited a smoother surface and a more uniform distribution of aggregates.
PubMed: 38824733
DOI: 10.1016/j.foodchem.2024.139903 -
Food Research International (Ottawa,... Jul 2024The effect of microfluidization treatment on the primary, secondary, and tertiary structure of soybean protein isolate (SPI) was investigated. The samples were treated...
The effect of microfluidization treatment on the primary, secondary, and tertiary structure of soybean protein isolate (SPI) was investigated. The samples were treated with and without controlling the temperature and circulated in the system 1, 3, and 5 times at high pressure (137 MPa). Then, the treated samples were freeze-dried and reconstituted in water to check the impact of the microfluidization on two different states: powder and solution. Regarding the primary structure, the SDS-PAGE analysis under reducing conditions showed that the protein bands remained unchanged when exposed to microfluidization treatment. When the temperature was controlled for the samples in their powder state, a significant decrease in the quantities of β-sheet and random coil and a slight reduction in α-helix content was noticed. The observed decrease in β-sheet and the increase in β-turns in treated samples indicated that microfluidization may lead to protein unfolding, opening the hydrophobic regions. Additionally, a lower amount of α-helix suggests a higher protein flexibility. After reconstitution in water, a significant difference was observed only in α-helix, β-sheet and β-turn. Related to the tertiary structure, microfluidization increases the surface hydrophobicity. Among all the conditions tested, the samples where the temperature is controlled seem the most suitable.
Topics: Soybean Proteins; Powders; Hydrophobic and Hydrophilic Interactions; Food Handling; Protein Structure, Secondary; Temperature; Pilot Projects; Electrophoresis, Polyacrylamide Gel; Glycine max; Solutions; Freeze Drying
PubMed: 38823863
DOI: 10.1016/j.foodres.2024.114466 -
Food Research International (Ottawa,... Jul 2024Myofibrillar proteins are crucial for gel formation in processed meat products such as sausages and meat patties. Freeze-thaw cycles can alter protein properties,...
Myofibrillar proteins are crucial for gel formation in processed meat products such as sausages and meat patties. Freeze-thaw cycles can alter protein properties, impacting gel stability and product quality. This study aims to investigate the potential of thawed drip and its membrane-separated components as potential antifreeze agents to retard denaturation, oxidation and gel deterioration of myofibrillar proteins during freezing-thawing cycles of pork patties. The thawed drip and its membrane-separated components of > 10 kDa and < 10 kDa, along with deionized water, were added to minced pork at 10 % mass fraction and subjected to increasing freeze-thaw cycles. Results showed that the addition of thawed drip and its membrane separation components inhibited denaturation and structural changes of myofibrillar proteins, evidenced by reduced surface hydrophobicity and carbonyl content, increased free sulfhydryl groups, protein solubility and α-helix, as compared to the deionized water group. Correspondingly, improved gel properties including water-holding capacity, textural parameters and denser network structure were observed with the addition of thawed drip and its membrane separation components. Denaturation and oxidation of myofibrillar proteins were positively correlated with gel deterioration during freezing-thawing cycles. We here propose a role of thawed drip and its membrane separation components as cryoprotectants against myofibrillar protein gel deterioration during freeze-thawing cycles.
Topics: Freezing; Animals; Gels; Swine; Muscle Proteins; Myofibrils; Food Handling; Protein Denaturation; Meat Products; Hydrophobic and Hydrophilic Interactions; Solubility; Water; Oxidation-Reduction
PubMed: 38823861
DOI: 10.1016/j.foodres.2024.114461 -
Nature Cell Biology Jun 2024
Topics: Humans; Endoplasmic Reticulum; Cytoplasmic Granules; Animals; Endoplasmic Reticulum Stress; Unfolded Protein Response
PubMed: 38822214
DOI: 10.1038/s41556-024-01434-7 -
International Journal of Biological... Jun 2024A simple but robust strategy of ball milling (20 Hz, 30 Hz for 30 s, 60 s, 120 s, 180 s) was utilized to modify bamboo shoots fiber (BSDF) in shrimp surimi. The...
A simple but robust strategy of ball milling (20 Hz, 30 Hz for 30 s, 60 s, 120 s, 180 s) was utilized to modify bamboo shoots fiber (BSDF) in shrimp surimi. The water holding capacity, swelling capacity, and oil binding capacity of 30 Hz-60 s milled BSDF exhibited the highest values of 5.61 g/g, 3.13 mL/g, and 6.93 g/g, significantly higher (P < 0.05) than untreated one (3.65 g/g, 2.03 mL/g, 4.57 g/g). Ball-milled BSDF exhibited a small-sized structure with the relative crystallinity decreased from 40.44 % (control) to 11.12 % (30 Hz-180 s). The myosin thermal stability, gelation properties of surimi were significantly enhanced by incorporating 20 Hz-120 s and 30 Hz-60 s BSDF via promoting protein unfolding, covalent hydrophobic interactions, and hydrogen bonding. A matrix-reinforcing and water entrapping effect was observed, exhibiting reinforced networks with down-sized water tunnels. However, BSDF modified at 180 s contributed to over-aggregated networks with fractures and enlarged gaps. Appropriate ball-milled BSDF (20 Hz-120 s, and 30 Hz-60 s) resulted in a significant decrease in α-helix (P < 0.05), accompanied by an increase of β-sheets and β-turn. This work could bring some insights into the applications of modified BSDF and its roles in the gelation of surimi-based food.
Topics: Animals; Dietary Fiber; Plant Shoots; Water; Chemical Phenomena; Myosins; Bambusa
PubMed: 38821812
DOI: 10.1016/j.ijbiomac.2024.131979 -
Biochemistry Jun 2024TDP-43 is a ubiquitously expressed, multidomain functional protein that is distinctively known to form aggregates in many fatal neurodegenerative disorders. However, the...
TDP-43 is a ubiquitously expressed, multidomain functional protein that is distinctively known to form aggregates in many fatal neurodegenerative disorders. However, the information for arresting TDP-43 aggregation is missing due to a lack of understanding of the molecular mechanism of the aggregation and structural properties of TDP-43. TDP-43 is inherently prone to aggregation and has minimal protein solubility. Multiple studies have been performed on the smaller parts of TDP-43 or the full-length protein attached to a large solubilization tag. However, the presence of co-solutes or solubilization tags is observed to interfere with the molecular properties and aggregation mechanism of full-length TDP-43. Notably, this study populated and characterized the native, dimeric state of TDP-43 without the interference of co-solutes or protein modifications. We observed that the electrostatics of the local environment is capable of the partial unfolding and monomerization of the native dimeric state of TDP-43 into an amyloidogenic molten globule. By employing the tools of thermodynamics and kinetics, we reveal the structural characteristics and temporal order of the early intermediates and transition states during the transition of the molten globule to β-rich, amyloid-like aggregates of TDP-43, which is governed by the electrostatics of the environment. The current advanced understanding of the nature of native and early aggregation-prone intermediates, early steps, and the influence of electrostatics in TDP-43 aggregation is essential for drug design.
Topics: DNA-Binding Proteins; Humans; Static Electricity; Protein Aggregates; Kinetics; Thermodynamics; Protein Multimerization; Amyloid; Amyloid beta-Protein Precursor
PubMed: 38820318
DOI: 10.1021/acs.biochem.4c00060 -
Analytical Chemistry Jun 2024Specific amino acid footprinting mass spectrometry (MS) is an increasingly utilized method for elucidating protein higher order structure (HOS). It does this by adding...
Specific amino acid footprinting mass spectrometry (MS) is an increasingly utilized method for elucidating protein higher order structure (HOS). It does this by adding to certain amino acid residues a mass tag, whose reaction extent depends on solvent accessibility and microenvironment of the protein. Unlike reactive free radicals and carbenes, these specific footprinters react slower than protein unfolding. Thus, their footprinting, under certain conditions, provokes structural changes to the protein, leading to labeling on non-native structures. It is critical to establish conditions (i.e., reagent concentrations, time of reaction) to ensure that the structure of the protein following footprinting remains native. Here, we compare the efficacy of five methods in assessing protein HOS following footprinting at the intact protein level and then further localize the perturbation at the peptide level. Three are MS-based methods that provide dose-response plot analysis, evaluation of Poisson distributions of precursor and products, and determination of the average number of modifications. These MS-based methods reliably and effectively indicate HOS perturbation at the intact protein level, whereas spectroscopic methods (circular dichroism (CD) and dynamic light scattering (DLS)) are less sensitive in monitoring subtle HOS perturbation caused by footprinting. Evaluation of HOS at the peptide level indicates regions that are sensitive to localized perturbations. Peptide-level analysis also provides higher resolution of the HOS perturbation, and we recommend using it for future footprinting studies. Overall, this work shows conclusive evidence for HOS perturbation caused by footprinting. Implementation of quality control workflows can identify conditions to avoid the perturbation, for footprinting, allowing accurate and reliable identification of protein structural changes that accompany, for example, ligand interactions, mutations, and changes in solution environment.
Topics: Proteins; Mass Spectrometry; Protein Footprinting; Protein Conformation; Amino Acids; Circular Dichroism
PubMed: 38815160
DOI: 10.1021/acs.analchem.4c01735 -
Progress in Molecular Biology and... 2024In order for an ordered protein to perform its specific function, it must have a specific molecular structure. Information about this structure is encoded in the... (Review)
Review
In order for an ordered protein to perform its specific function, it must have a specific molecular structure. Information about this structure is encoded in the protein's amino acid sequence. The unique functional state is achieved as a result of a specific process, known as protein folding. However, as a result of partial or complete unfolding of the polypeptide chain, proteins may misfold and aggregate, leading to the formation of various aggregated structures, such as like amyloid aggregates with the cross-β structure. A variety of cellular biological processes can be affected by protein aggregates that consume essential factors necessary for maintaining proteostasis, which leads to the proteostasis imbalance and further accumulation of protein aggregates, often resulting in age-related neurodegenerative disease progression and aging. However, in addition to their well-established pathological effects, amyloids also play various physiological roles, and many important biological processes involve such 'functional amyloids'. This chapter represents a brief overview of the protein aggregation phenomenon outlines a timeline provides of some key discoveries in this exciting field.
Topics: Humans; Protein Aggregates; Animals; Amyloid; Protein Aggregation, Pathological; Protein Folding; Proteins
PubMed: 38811077
DOI: 10.1016/bs.pmbts.2024.03.007 -
Molecular Biology of the Cell Jul 2024undergo age-dependent declines in muscle organization and function, similar to human sarcopenia. The chaperone UNC-45 is required to fold myosin heads after translation...
undergo age-dependent declines in muscle organization and function, similar to human sarcopenia. The chaperone UNC-45 is required to fold myosin heads after translation and is likely used for refolding after thermally- or chemically-induced unfolding. UNC-45's TPR region binds HSP-90 and its UCS domain binds myosin heads. We observe early onset sarcopenia when UNC-45 is reduced at the beginning of adulthood. There is sequential decline of HSP-90, UNC-45, and MHC B myosin. A mutation in delays sarcopenia and loss of HSP-90, UNC-45, and myosin. UNC-45 undergoes age-dependent phosphorylation, and mass spectrometry reveals phosphorylation of six serines and two threonines, seven of which occur in the UCS domain. Additional expression of UNC-45 results in maintenance of MHC B myosin and suppression of A-band disorganization in old animals. Our results suggest that increased expression or activity of UNC-45 might be a strategy for prevention or treatment of sarcopenia.
Topics: Animals; Caenorhabditis elegans Proteins; Caenorhabditis elegans; Aging; Molecular Chaperones; Myosins; Sarcomeres; Phosphorylation; HSP90 Heat-Shock Proteins; Humans; Mutation; Muscle, Skeletal
PubMed: 38809582
DOI: 10.1091/mbc.E23-12-0488 -
Protein Science : a Publication of the... Jun 2024In eukaryotes, the ubiquitin-proteasome system is responsible for intracellular protein degradation. Proteins tagged with ubiquitin are recognized by ubiquitin receptors...
In eukaryotes, the ubiquitin-proteasome system is responsible for intracellular protein degradation. Proteins tagged with ubiquitin are recognized by ubiquitin receptors on the 19S regulatory particle (RP) of the 26S proteasome, unfolded, routed through the translocation channel of the RP, and are then degraded in the 20S core particle (CP). Aromatic paddles on the pore-1 loops of the RP's Rpt subunits grip the substrate and pull folded domains into the channel, thereby unfolding them. The sequence that the aromatic paddles grip while unfolding a substrate is therefore expected to influence the extent of unfolding, and low complexity sequences have been shown to interfere with grip. However, the detailed spatial requirements for grip while unfolding proteins, particularly from the N-terminus, remain unknown. We determined how the location of glycine-rich tracts relative to a folded domain impairs unfolding. We find that, in contrast to a previous report, inserting glycine-rich sequences closer to the folded domain reduced unfolding ability more than positioning them further away. Locations that have the biggest effect on unfolding map onto the regions where the aromatic paddles are predicted to interact with the substrate. Effects on unfolding from locations up to 67 amino acids away from the folded domain suggest that there are additional interactions between the substrate and the proteasome beyond the aromatic paddles that facilitate translocation of the substrate. In sum, this study deepens understanding of the mechanical interactions within the substrate channel by mapping the spacing of interactions between the substrate and the proteasome during unfolding.
Topics: Proteasome Endopeptidase Complex; Models, Molecular; Humans; Protein Unfolding; Protein Transport
PubMed: 38801231
DOI: 10.1002/pro.5034