Loss Of Tryptophan Research Articles

Antioxidant Properties of Platycladus orientalis Flavonoids for Treating UV-Induced Damage in Androgenetic Alopecia Hair.

Androgenetic alopecia (AGA) causes thinning hair, but poor hair quality in balding areas and damage from UV radiation have been overlooked. Plant extracts like Platycladus orientalis flavonoids (POFs) may improve hair quality in AGA. This study examines POFs' effectiveness in treating AGA-affected hair and repairing UV-induced damage. Hair samples were analyzed using scanning electron microscopy (SEM) to examine surface characteristics, electron paramagnetic resonance (EPR) spectroscopy to measure free radicals in the hair, and spectrophotometry to assess changes in hair properties. POFs effectively removed hydroxyl radicals from keratinocytes and had antioxidant properties. They also reduced UV-induced damage to AGA hair by mitigating the production of melanin free radicals. Following POF treatment, the reduction in peroxidized lipid loss in AGA hair was notable at 59.72%, thereby effectively delaying the progression of hair color change. Moreover, protein loss decreased by 191.1 μ/g and tryptophan loss by 15.03%, ultimately enhancing hair's tensile strength. compared to healthy hair, hair damaged by AGA shows more pronounced signs of damage when exposed to UV radiation. POFs help protect balding hair by reducing oxidative damage and slowing down melanin degradation.

Dietary tryptophan deficiency promotes gut RORγt+ Treg cells at the expense of Gata3+ Treg cells and alters commensal microbiota metabolism

Micronutrient deficiency is a major cause of disease throughout the world. Yet, how perturbations influence the immune-microbiome interface remains poorly understood. Here, we report that loss of dietary tryptophan (Trp) reshapes intestinal microbial communities, including the depletion of probiotic L.reuteri, drives transcriptional changes to immune response genes in the intestinal ileum, and reshapes the regulatory Tcell (Treg) compartment. Dietary Trp deficiency promotes expansion of RORγt+ Treg cells and the loss of Gata3+ Tregs in a microbiota-dependent manner. In the absence of dietary Trp, provision of the AhR ligand indole-3-carbinol is sufficient to restore the Treg compartment. Together, these data show that dietary Trp deficiency perturbs the interaction between the host and its bacterial symbionts to regulate Treg homeostasis via the deprivation of bacterially derived Trp metabolites. Our findings highlight an essential role for immune-microbiome crosstalk as a key homeostatic regulator during nutrient deficiency.

Influence of plasma halide, pseudohalide and nitrite ions on myeloperoxidase-mediated protein and extracellular matrix damage

Myeloperoxidase (MPO) mediates pathogen destruction by generating the bactericidal oxidant hypochlorous acid (HOCl). Formation of this oxidant is however associated with host tissue damage and disease. MPO also utilizes H2O2 to oxidize other substrates, and we hypothesized that mixtures of other plasma anions, including bromide (Br−), iodide (I−), thiocyanate (SCN−) and nitrite (NO2−), at normal or supplemented concentrations, might modulate MPO-mediated HOCl damage. For the (pseudo)halide anions, only SCN− significantly modulated HOCl formation (IC50 ∼33 μM), which is within the normal physiological range, as judged by damage to human plasma fibronectin or extracellular matrix preparations detected by ELISA and LC-MS. NO2− modulated HOCl-mediated damage, in a dose-dependent manner, at physiologically-attainable anion concentrations. However, this was accompanied by increased tyrosine and tryptophan nitration (detected by ELISA and LC-MS), and the overall extent of damage remained approximately constant. Increasing NO2− concentrations (0.5–20 μM) diminished HOCl-mediated modification of tyrosine and methionine, whereas tryptophan loss was enhanced. At higher NO2− concentrations, enhanced tyrosine and methionine loss was detected. These analytical data were confirmed in studies of cell adhesion and metabolic activity. Together, these data indicate that endogenous plasma levels of SCN− (but not Br− or I−) can modulate protein modification induced by MPO, including the extent of chlorination. In contrast, NO2− alters the type of modification, but does not markedly decrease its extent, with chlorination replaced by nitration. These data also indicate that MPO could be a major source of nitration in vivo, and particularly at inflammatory sites where NO2− levels are often elevated.

Bioactive β-Carbolines Harman and Norharman in Sesame Seed Oils in China.

The β-carbolines in our diet, mainly including harman and norharman, are a group of biologically active, naturally occurring plant-derived alkaloids. Fragrant sesame seed oil is one of the most popular flavor edible oils in China. Considering that sesame seeds are roasted at 200–240 °C during the processing of flavor sesame seed oils, it is meaningful to investigate the levels of β-carboline compounds in various sesame seed oils. In this work, the levels of β-carbolines (harman and norharman) in different types of sesame seed oils in China (e.g., pressed fragrant sesame oil, ground fragrant sesame oil) have been determined systematically. The results showed that the levels of total β-carbolines in pressed fragrant sesame oils (700.5~2423.2 μg/kg) were higher than that in ground fragrant sesame oils (660.4~1171.7 μg/kg). Roasting sesame seeds at high temperatures (200–240 °C) led to higher levels of β-carbolines (660~2400 μg/kg) in fragrant sesame seed oils. In addition, the loss of tryptophan might be attributed to the formation of β-carbolines in sesame seeds during the roasting process. In general, fragrant sesame seed oils (pressed fragrant sesame oils, ground fragrant sesame oils) contain higher levels of β-carbolines due to the formation of harman and norharman during the roasting sesame seed process.

Novel NPM1 exon 5 mutations and gene fusions leading to aberrant cytoplasmic nucleophosmin in AML

AbstractNucleophosmin (NPM1) mutations in acute myeloid leukemia (AML) affect exon 12, but also sporadically affect exons 9 and 11, causing changes at the protein C-terminal end (tryptophan loss, nuclear export signal [NES] motif creation) that lead to aberrant cytoplasmic NPM1 (NPM1c+), detectable by immunohistochemistry. Combining immunohistochemistry and molecular analyses in 929 patients with AML, we found non–exon 12 NPM1 mutations in 5 (1.3%) of 387 NPM1c+ cases. Besides mutations in exons 9 (n = 1) and 11 (n = 1), novel exon 5 mutations were discovered (n = 3). Another exon 5 mutation was identified in an additional 141 patients with AML selected for wild-type NPM1 exon 12. Three NPM1 rearrangements (NPM1/RPP30, NPM1/SETBP1, NPM1/CCDC28A) were detected and characterized among 13 979 AML samples screened by cytogenetic/fluorescence in situ hybridization and RNA sequencing. Functional studies demonstrated that in AML cases, new NPM1 proteins harbored an efficient extra NES, either newly created or already present in the fusion partner, ensuring its cytoplasmic accumulation. Our findings support NPM1 cytoplasmic relocation as critical for leukemogenesis and reinforce the role of immunohistochemistry in predicting AML-associated NPM1 genetic lesions. This study highlights the need to develop new assays for molecular diagnosis and monitoring of NPM1-mutated AML.

Two Novel Mutations of the NPM1 Gene in Syrian Adult Patients with Acute Myeloid Leukemia and Normal Karyotype.

Objective:Somatic mutations in exon 12 of the NPM1 gene is one of the most common genetic abnormalities in adult acute myeloid leukemia (AML), which is observed in 25-35% of AML patients and in 50-60% of patients with cytogenetically normal AML (CN-AML). Methods:We performed Sanger sequencing of exon 12 of the NPM1 gene, on 44 CN-AML patients to characterize NPM1 status. Results:In this study, NPM1 mutations were identified in 10 (22.7%) of the 44 CN-AML patients. Among the 10 patients with NPM1 mutations, type A NPM1 mutations were identified in 8 (80%) patients, whereas non-A type NPM1 mutations were observed in 2 (20%) patients. Two non-A type NPM1 mutations were not previously reported: c.867-868InsCGGA and c.861-862InsTGCA. These two novel mutant proteins display a nuclear export signal (NES) motif (L-xxx-L-xx-V-x-L) less frequently and L-x-Lx-V-xx-V-x-L it has been never seen before, yet. However, both novel mutations show a tryptophan loss at codon 288 and 290 at the mutant C-terminus which are crucial for aberrant nuclear export of NPM into the cytoplasm. Conclusions:This study suggests previously unreported NPM1 mutations may be non-rare and thus additional sequence analysis is needed along with conventional targeted mutational analysis to detect non type-A NPM1 mutations.

Lipid oxidation and protein co-oxidation in ready-to-eat meat products as affected by temperature, antioxidant, and packaging material during 6 months of storage.

Ambient-storage-friendly, ready-to-eat (RTE) meat products are convenient in emergencies, such as earthquakes, flash floods and the current global Covid-19 lockdown. However, given the processing and long storage time of such food products, the lipid and protein components may be more susceptible to oxidation. Chicken serunding is a low-moisture, high-lipid, high-protein, RTE product that is prone to lipid oxidation and protein co-oxidation, causing product quality deterioration. The present study assessed the effects of storage temperature (25, 40, 60 °C), antioxidant (butylated hydroxyanisole, BHA), and multilayer packaging materials [metallised polyethene terephthalate (MPET) and aluminium] on the lipid oxidation and protein co-oxidation of chicken serunding during six months of storage. All lipid and protein markers elevated with increasing temperature (25 < 40 < 60 °C), indicating that storage of low-moisture meat at high temperature is not feasible. BHA was effective against lipid oxidation, as indicated by the significantly lower (p <0.05) extracted lipid content and delayed formation of malondialdehyde, a secondary lipid oxidation product. However, BHA is not effective against protein co-oxidation, as shown by the insignificant (p >0.05) effect on preventing tryptophan loss, protein carbonyl formation and Schiff base accumulation. MPET packaging with a superior light and oxygen barrier provided significant protection (p <0.05) compared to aluminium. In conclusion, low temperature (25 °C) storage of low-moisture, high-lipid, high-protein, cooked meat systems in MPET packaging is recommended for long-term storage to delay the progression of lipid oxidation and protein co-oxidation.

Effect of different extent of protein oxidation on the frozen storage stability of muscle protein in obscure pufferfish (Takifugu obscurus)

The objective of this study was to explore frozen storage stability of muscle protein of the obscure pufferfish using a semi-model system based on light exposure (LE), oxygen concentration (OC), and multiple freeze-thaw cycles (FTCs). The dorsal muscle of fifteen frozen fish was cut into small pieces (2 cm × 1.5 cm × 1 cm) and divided into 4 groups stored in differing conditions: one was stored in light with 0% oxygen (L-0), one in light with 20% oxygen (L-20), one in dark with 0% oxygen (D-0), and one in dark with 20% oxygen (D-20). All were subsequently subjected to five FTCs. Protein oxidation (PO) levels, protein structure, and water states were all assessed. The results showed that both LE and OC could significantly accelerate sulfhydryl and tryptophan loss, in the order of L-20 > L-0 and D-20 > D-0 during FTCs. Changes of protein structure were in line with the extent of PO indicated by surface hydrophobicity and UV absorbance. An increased PO also affected the water states, with higher PO resulting in immobilized water being more tightly trapped within the protein. However, thawing loss was shown to be independent of PO and the structural changes of myofibrillar protein.

Preparation of horsegram protein concentrate with improved protein quality, in vitro digestibility and available lysine

Horsegram (Macrotyloma uniflorum or Dolichos biflorus), an inexpensive pulse crop with high lysine and iron contents, is underutilized due to the presence of antinutrients like lectin and trypsin inhibitors, which limit protein digestibility and availability. Horsegram protein concentrate (HGPC) was prepared, which had 80.4 ± 3.5% protein and 94.2 ± 1.4% in vitro protein digestibility compared to dehulled horsegram flour (22.8 ± 0.8% and 82.3 ± 1.2%, respectively). Available lysine content in concentrate was increased by 64% compared to dehulled horsegram flour. The trypsin inhibitory activity in the protein concentrate decreased by 51% from 36.6 ± 3.5 TIU/mg in horsegram flour to 17.6 ± 2.5 TIU/mg in HGPC. Nutritional indices, including PDCAAS values for different age groups, were calculated and found to be slightly lower due to the loss of methionine and tryptophan in the concentrate. However, branched amino acids and lysine contents were higher. Thus, a vegetarian source with high protein digestibility and available lysine content could be prepared as a protein ingredient for the food industry.

Investigation of the effect of sodium nitrite on protein oxidation markers in food protein suspensions.

The aim of this study is to investigate the effect of sodium nitrite (NaNO2 ) on protein oxidation and the use of 3-nitrotyrosine (3NT) as a protein oxidation marker in suspensions of the food protein. Food proteins, namely bovine serum albumin, casein, and myofibrillar protein, were suspended in 100mM sodium phosphate buffer and nitrated with 25µM iron (III) chloride, 2.5mM hydrogen peroxide, and 150mg/kg NaNO2 at 37°C for a period of 24hr. The food protein suspensions were analyzed at different sampling periods for the loss of tryptophan (TRY) residues as well as the formation of Schiff bases (SBs), protein carbonyls, 3NT, and dityrosine (DT). It was found that NaNO2 has pro-oxidant activity in NaNO2 -added food protein suspensions due to the increased amounts of SBs, protein carbonyls, 3NT, and DT as well as decreased TRY fluorescence. Positive correlations between the 3NT and other protein oxidation markers except for TRY fluorescence were found in NaNO2 -added food protein suspensions. In conclusion, these findings on the detection of 3NT indicate that it might be a useful tool as a new protein oxidation biomarker in food samples. PRACTICAL APPLICATIONS: Sodium nitrite (NaNO2 ) supports protein oxidation in different food protein suspension. 3-nitrotyrosine (3NT) was found in food protein suspensions and can be a potential biomarker for protein nitration in food sample due to potential relationship between 3NT and other oxidation markers. The results showed that this study has formed novel insight into interaction between NaNO2 and food proteins and indicate that it might affect the food quality and its nutritional value. Moreover, the relationship between protein oxidation/nitration and food quality as well as the comprehension of the scientific and technological meaning of these phenomena has been hindered because of the lack of knowledge about the basic chemistry behind the protein oxidation and nitration pathways. For the clarification of these phenomena, further studies are still needed.

Assembling of the interfacial layer affects the physical and oxidative stability of faba bean protein-stabilized oil-in-water emulsions with chitosan

This study investigated how different ways of assembling of faba bean protein isolate (FBPI) and chitosan (CH) affected the oil-in-water emulsion stability. Emulsions were prepared either by a soluble FBPI-CH complex to make EMC, or by FBPI first and then added non-interactive CH to make EMN, or by FBPI first and then added a secondary layer of CH via electrostatic attraction to make EML. A control emulsion EMFB was prepared by only FBPI. All emulsions contained 5% (w/v) oil and 0.5% (w/v) FBPI. EMC/EML/EMN contained 0.05% (w/v) CH. The emulsion stability was monitored at 37 °C for 7 days. EML not only showed better physical stability as indicated by the changes in droplet size, Turbiscan stability index, and microstructure (by confocal microscopy), but also exhibited superior oxidative stability as indicated by lipid oxidation products (less conjugated dienes and secondary oxidation products) and protein oxidation biomarkers (less tryptophan loss and carbonyls formation). This might be attributed to that CH enhanced formation of a layer-by-layer (LBL) interfacial structure, which increased the interfacial layer thickness/compactness and maintained the interfacial protein adsorption. In contrast, EMC demonstrated worse stability, albeit with thicker interfacial layer due to CH. This could be explained by the lower surface hydrophobicity and surface charge of soluble FBPI-CH complex. The free aqueous CH also impaired physical stability and promoted oxidation, as shown in EMN. In conclusion, CH improves the oxidative and physical stability of FBPI-containing emulsion when added as a LBL model.

Tryptophan Intake and Tryptophan Losses in Hemodialysis Patients: A Balance Study.

Tryptophan depletion is common in hemodialysis patients. The cause of this depletion remains largely unknown, but reduced nutritional tryptophan intake, losses during dialysis or an increased catabolism due to an inflammatory state are likely contributors. Currently, little is known about tryptophan homeostasis in hemodialysis patients. We assessed dietary tryptophan intake, measured plasma tryptophan during dialysis, and measured the combined urinary and dialysate excretion of tryptophan in 40 hemodialysis patients (66 ± 15 years and 68% male). Patients had low tryptophan concentrations (27 ± 9 µmol/L) before dialysis. Mean dietary tryptophan intake was 4454 ± 1149 µmol/24 h. Mean urinary tryptophan excretion was 15.0 ± 12.3 μmol/24 h, dialysate excretion was 209 ± 67 μmol/24 h and combined excretion was 219 ± 66 µmol/24 h, indicating only 5% of dietary tryptophan intake was excreted. No associations were found between plasma tryptophan concentration and tryptophan intake, plasma kynurenine/tryptophan ratio or inflammatory markers. During dialysis, mean plasma tryptophan concentration increased 16% to 31 ± 8 µmol/L. Intradialytic increase in plasma tryptophan was associated with a lower risk of mortality, independent of age, sex and dialysis vintage (HR: 0.87 [0.76–0.99]; P = 0.04). Tryptophan intake was well above the dietary recommendations and, although tryptophan was removed during dialysis, mean plasma tryptophan increased during dialysis. The cause of this phenomenon is unknown, but it appears to be protective.

Effects of Storage Time and Temperature on Lipid Oxidation and Protein Co-Oxidation of Low-Moisture Shredded Meat Products.

Studies on the oxidative changes in meat-based, low-moisture, ready to eat foods are complicated due to complex food system and slow lipid-protein oxidative deterioration. The current study evaluates the oxidative changes over six months of storage on shredded beef and chicken products (locally known as serunding) for physicochemical analysis, lipid oxidation (conjugated dienes and malondialdehydes) and protein co-oxidation (soluble protein content, amino acid composition, protein carbonyl, tryptophan loss and Schiff base fluorescence) at 25 °C, 40 °C and 60 °C. The lipid stability of chicken serunding was significantly lower than beef serunding, illustrated by higher conjugated dienes content and higher rate of malondialdehyde formation during storage. In terms of protein co-oxidation, chicken serunding with higher polyunsaturated fatty acids (PUFA) experienced more severe oxidation, as seen from lower protein solubility, higher protein carbonyl and Schiff base formation compared to beef serunding. To conclude, chicken serunding demonstrates lower lipid and protein stability and exhibits higher rate of lipid oxidation and protein co-oxidation than beef serunding. These findings provide insights on the progression of lipid oxidation and protein co-oxidation in cooked, shredded meat products and could be extrapolated to minimize possible adverse effects arising from lipid oxidation and protein co-oxidation, on the quality of low-moisture, high-lipid, high-protein foods.

An allosteric MALT1 inhibitor is a molecular corrector rescuing function in an immunodeficient patient.

MALT1 paracaspase is central for lymphocyte antigen-dependent responses including NF-κB activation. We discovered nanomolar, selective allosteric inhibitors of MALT1 that bind by displacing the side chain of Trp580, locking the protease in an inactive conformation. Interestingly, we had previously identified a patient homozygous for a MALT1 Trp580-to-serine mutation who suffered from combined immunodeficiency. We show that the loss of tryptophan weakened interactions between the paracaspase and C-terminal immunoglobulin MALT1 domains resulting in protein instability, reduced protein levels and functions. Upon binding of allosteric inhibitors of increasing potency, we found proportionate increased stabilization of MALT1-W580S to reach that of wild-type MALT1. With restored levels of stable MALT1 protein, the most potent of the allosteric inhibitors rescued NF-κB and JNK signaling in patient lymphocytes. Following compound washout, MALT1 substrate cleavage was partly recovered. Thus, a molecular corrector rescues an enzyme deficiency by substituting for the mutated residue, inspiring new potential precision therapies to increase mutant enzyme activity in other deficiencies.

Blue Death of Nematodes

This paper shows that the aging and death of nematodes, accompanied by the ignition of a blue glow under fluorescent microscopy, are not directly linked to any lipofuscin (aging pigment), nor with the anthranilic acid (a product of degradation of tryptophan residues of proteins). The main contribution in the blue flash of the dying nematodes belongs to parasitic light, scattered on the cuticle and bodies of the worm. The main contribution in the blue region at spectrofluorometry of homogenates, obtained from nematodes, really gives anthranilic acid. However, the content of anthranilic acid, measured by spectrofluorimetry, in adult nematodes is lower than that in the young ones. Artificial aging of nematodes by moderate heating revealed no accumulation of anthranilate and no loss of tryptophan, from which it must be formed. Thus, it is hardly lipofuscin or anthranilic acid. The cause of aging and death of nematodes is the formation of strong cross-links between proteins. This is supported by data on tryptophan fluorescence and light scattering of homogenates: the old worms show a large number of denaturated proteins and large protein particles with a strong cross-links, which are not destroyed be detergent.

Lonicera japonica Thunb. extract improves the quality of cold‐stored porcine patty through inhibition of lipid and myofibrillar protein oxidation

SummaryThe aim of this work was to analyse the honeysuckle extract (HE) for its in vitro antioxidant activity using the ORAC, reducing power, DPPH and ABTS assays. The effects of HE on the quality characteristics and physicochemical properties of porcine patties during refrigerated storage at 4 °C were studied. Chlorogenic acid contributed about 50% to the antioxidant activity of HE. HE possessing strong antioxidant activity was incorporated into porcine patties at 0.02%, 0.1% and 0.5% concentrations and compared with 0.02% and 0.1% chlorogenic acid as references. The addition of 0.1% HE and chlorogenic acid significantly reduced the loss of hardness, springiness, chewiness and cohesiveness of porcine patties in terms of inhibition of lipid oxidation, formation of protein carbonyls, generation of covalent bonds and tryptophan loss during 15 days of chilled storage. HE could be used in porcine patty as a natural antioxidant.

Entrapment of amino acids in gas phase surfactant assemblies: The case of tryptophan confined in positively charged (1R,2S)-dodecyl (2-hydroxy-1-methyl-2-phenylethyl) dimethylammonium bromide aggregates.

The ability of positively charged aggregates of the surfactant (1R,2S)-dodecyl(2-hydroxy-1-methyl-2-phenylethyl)dimethylammonium bromide (DMEB) to incorporate D-tryptophan or L-tryptophan in the gas phase has been investigated by electrospray ion mobility mass spectrometry (ESI-IM-MS). Strongly impacted by the pH of the electrosprayed solutions, both protonated (T+ ) and deprotonated (T- ) tryptophan are effectively included into the aggregates, whereas, tryptophan in zwitterionic (T0 ) form is practically absent in singly charged DMEB aggregates but can be found in multiply charged ones. The ability to incorporate tryptophan increases with the aggregation number and charge state of aggregates. More than 1 tryptophan species can be entrapped (aggregates including up to 5 tryptophan are observed). Collision induced dissociation experiments performed on the positively singly charged DMEB hexamer containing 1 T- show that at low collision energies the loss of a DMEB molecule is preferred with respect to the loss of the DMEB cation plus T- species which, in turn, is preferred with respect to the loss of mere tryptophan, suggesting that the deprotonated amino acid is preferentially located in proximity of a DMEB head group and with the ionic moiety pointing towards the core of the aggregate. The analysis of the collision cross sections (CCS) of bare and tryptophan containing aggregates allowed evaluating the contributions of tryptophan and bromide ions to the total aggregate CCS. No significant discrimination between D-tryptophan and L-tryptophan by the chiral DMEB aggregates has been evidenced by mass spectra data, CID experiments, and CCS values.

Chlorogenic acid induced colored reactions and their effect on carbonyls, phenolic content, and antioxidant capacity in sunflower butter cookies

The high chlorogenic acid (CGA) content of sunflower seeds causes a greening reaction in sunflower butter baked products which can deter application of sunflower butter as an allergen-free alternative to other plant and dairy based butters. This study focused on how greening intensity of sunflower butter cookies made with different sweeteners (maple, agave, corn syrups, honey and xylitol) affected greening, protein oxidation products, Folin and ABTS•+ radical scavenging ability. Cookies made with maple syrup and xylitol had higher pH and resulted in more greening. The dough made with agave syrup had highest total carbonyls caused by its highest reducing sugar content resulting in more Maillard reaction during dough preparation, while after baking cookies with highest greening (maple syrup) and highest reducing sugar (agave syrup) had higher carbonyls than other sweetener treatments. Cookies made with maple syrup and xylitol also had lower folin-ciocalteau reagent reducing capacity and tryptophan fluorescence. The greening reaction did not affect Schiff bases from oxidation and antioxidant capacity in cookies made with different sweeteners. Higher pH sweeteners thus enhanced greening intensity, tryptophan loss and lowered the total phenolic content after baking and storage, but did not influence the ABTS•+ capacity of sunflower butter cookies.

Oxidative damage to food and human serum proteins: Radical-mediated oxidation vs. glyco-oxidation

This study compared a hydroxyl radical-generating system (HRGS) (0.05–0.2mM Fe3++0.6mM H2O2) and a glycation system (GLY) (0.05–0.2mM Fe3++0.05M glucose) for their ability to promote protein carbonylation and tryptophan depletion in myofibrillar proteins, ovalbumin, β-lactoglobulin, soy protein and human serum albumin. Animal-source were more susceptible to protein carbonylation than soy proteins and globular were more susceptible than fibrillar proteins. Both systems promoted tryptophan loss and the formation of protein carbonyls and iron had a clear dose-effect in most systems and proteins. In the tested conditions, the GLY environment was more effective than the HRGS system in promoting the oxidative damage to food proteins. According to the results, glucose and H2O2 may compete for iron for the production of glycosylative and oxidative species, respectively. This study provides original insight into the chemical mechanisms implicated in the oxidative and glycosylative damage to food proteins.

Key role of cysteine residues and sulfenic acids in thermal- and H2O2-mediated modification of β-lactoglobulin

Oxidation results in protein deterioration in mammals, plants, foodstuffs and pharmaceuticals, via changes in amino acid composition, fragmentation, aggregation, solubility, hydrophobicity, conformation, function and susceptibility to digestion. This study investigated whether and how individual or combined treatment with heat, a commonly encountered factor in industrial processing, and H2O2 alters the structure and composition of the major whey protein β-lactoglobulin. Thermal treatment induced reducible cross-links, with this being enhanced by low H2O2 concentrations, but decreased by high concentrations, where fragmentation was detected. Cross-linking was prevented when the single free Cys121 residue was blocked with iodoacetamide. Low concentrations of H2O2 added before heating depleted thiols, with H2O2 alone, or H2O2 added after heating, having lesser effects. A similar pattern was detected for methionine loss and methionine sulfoxide formation. Tryptophan loss was only detected with high levels of H2O2, and no other amino acid was affected, indicating that sulfur-centered amino acids are critical targets. No protection against aggregation was provided by high concentrations of the radical scavenger 5, 5-dimethyl-1-pyrroline N-oxide (DMPO), consistent with molecular oxidation, rather than radical reactions, being the major process. Sulfenic acid formation was detected by Western blotting and LC–MS/MS peptide mass-mapping of dimedone-treated protein, consistent with these species being significant intermediates in heat-induced cross-linking, especially in the presence of H2O2. Studies using circular dichroism and intrinsic fluorescence indicate that H2O2 increases unfolding during heating. These mechanistic insights provide potential strategies for modulating the extent of modification of proteins exposed to thermal and oxidant treatment.