Proteomics of Muscle-Specific Beef Color Stability Joseph, Poulson; Suman, Surendranath P; Rentfrow, Gregg ...
Journal of agricultural and food chemistry,
03/2012, Letnik:
60, Številka:
12
Journal Article
Recenzirano
The objective of the present study was to differentiate the sarcoplasmic proteome of color-stable (Longissimus lumborum; LL) and color-labile (Psoas major; PM) beef muscles. LL and PM muscles from ...seven beef carcasses (24 h post-mortem) were fabricated into 2.54 cm steaks, aerobically packaged, and assigned to refrigerated retail display for 9 days. LL steaks demonstrated greater (P < 0.05) color stability and lower (P < 0.05) lipid oxidation than PM steaks. Proteome analyses identified 16 differentially abundant proteins in LL and PM, including antioxidant proteins and chaperones. Proteins demonstrating positive correlation with redness (aldose reductase, creatine kinase, and β-enolase) and color stability (peroxiredoxin-2, peptide methionine sulfoxide reductase, and heat shock protein-27 kDa) were overabundant in LL, whereas the protein overabundant in PM (mitochondrial aconitase) exhibited negative correlation with redness. The color stability of LL could be attributed to the overabundance of antioxidant proteins and chaperones, and this finding suggests the necessity of developing muscle-specific processing strategies to improve beef color.
Lactate dehydrogenase (LDH) activity can regenerate NADH, which is a critical component in metmyoglobin reduction. However, limited research has determined the effects of lipid oxidation products on ...LDH activity. The overall objective of this study was to determine the effects of 4-hydroxy-2-nonenal (HNE) on LDH activity. LDH was reacted with HNE at pH 5.6 and 7.4, and LDH activity was measured as NADH formation following the addition of lactate and NAD. The effects of HNE on NADH-dependent metmyoglobin reduction also were analyzed. Mass spectrometric examination revealed that HNE adducts to LDH at both pH 5.6 and 7.4. More specifically, HNE binds with cysteine and histidine residues of LDH at pH 5.6 and 7.4. Covalent binding of HNE decreased NADH formation and metmyoglobin reduction (P < 0.05). These results indicate that secondary lipid oxidation products can inactivate enzymes involved in metmyoglobin reduction and have the potential to increase beef discoloration.
The objective of the present study was to characterize the proteome basis for intramuscular color stability variations in beef semimembranosus. Semimembranosus muscles from eight carcasses (n=8) were ...fabricated into 2.54-cm thick color-labile inside (ISM) and color-stable outside (OSM) steaks. One steak for sarcoplasmic proteome analysis was immediately frozen, whereas other steaks were allotted to retail display under aerobic packaging. Color attributes were evaluated instrumentally and biochemically on 0, 2, and 4days. Sarcoplasmic proteome was analyzed using two-dimensional electrophoresis and tandem mass spectrometry. ISM steaks demonstrated greater (P<0.01) abundance of glycolytic enzymes (fructose-bisphosphate aldolase A, phosphoglycerate mutase 2, and beta-enolase) and phosphatidylethanolamine-binding protein 1 than their OSM counterparts. Possible rapid post-mortem glycolysis in ISM, insinuated by over-abundance of glycolytic enzymes, could lead to rapid pH decline during early post-mortem, which in turn could potentially compromise its color stability. These results indicated that differential abundance of sarcoplasmic proteome contributes to intramuscular variations in beef color stability.
•Inside and outside beef semimembranosus regions exhibit color stability variations.•Proteome basis for the intramuscular variation in color stability was examined.•Glycolytic enzymes were over-abundant in color-labile inside semimembranosus.•Differences in sarcoplasmic proteome contribute to intramuscular variations.
The sarcoplasmic proteome of beef Longissimus lumborum demonstrating animal-to-animal variation in color stability was examined to correlate proteome profile with color. Longissimus lumborum (36h ...post-mortem) muscles were obtained from 73 beef carcasses, aged for 13days, and fabricated to 2.5-cm steaks. One steak was allotted to retail display, and another was immediately vacuum packaged and frozen at −80°C. Aerobically packaged steaks were stored under display, and color was evaluated on days 0 and 11. The steaks were ranked based on redness and color stability on day 11, and ten color-stable and ten color-labile carcasses were identified. Sarcoplasmic proteome of frozen steaks from the selected carcasses was analyzed. Nine proteins were differentially abundant in color-stable and color-labile steaks. Three glycolytic enzymes (phosphoglucomutase-1, glyceraldehyde-3-phosphate dehydrogenase, and pyruvate kinase M2) were over-abundant in color-stable steaks and positively correlated (P<0.05) to redness and color stability. These results indicated that animal variations in proteome contribute to differences in beef color.
•Proteome basis of animal-to-animal variation in beef color stability was examined.•Glycolytic enzymes were over-abundant in color-stable Longissimus lumborum steaks.•Three glycolytic enzymes were positively correlated to redness and color stability.•Differences in sarcoplasmic proteome contribute to beef color stability variations.
Pale, Soft, and Exudative (PSE) broiler breast meat has poor protein functionality, which leads to quality problems and economic loss in the poultry industry. Proteomics has been applied to ...characterize the biochemical mechanisms governing tenderness, color, and water-holding capacity in meat. However, the proteome basis of PSE has not yet been characterized for broiler breast meat. Therefore, this study was conducted to determine the differences in meat quality (cooking loss and shear force), descriptive sensory characteristics, consumer acceptance, and whole muscle proteome between normal and PSE-like broiler breast meat. Male Hubbard × Cobb 500 birds (n = 1,050) were raised in commercial houses. Prior to harvest, a sample of the broilers (n = 900) were subjected to short-term stress (38°C for 2 h), and the remaining broilers (n = 150) were maintained at control conditions (21°C for 2 h). Broiler breast (Pectoralis major) meat was collected and characterized by pH24 and L*24 as normal (pH24 5.8 to 6.2, L*24 45 to 55) or PSE-like (pH24 5.4 to 5.7, L*
24 55 to 65) samples. Normal broiler breast meat had lower shear force values than PSE-like meat (P < 0.05). Based on sensory descriptive analysis, normal cooked chicken breast meat was more tender and juicier than PSE-like breast meat (P < 0.05). Consumer sensory analysis results indicated that 81% of consumer panelists liked normal breast meat whereas 62% of the panelists liked PSE-like breast meat. Whole muscle proteome profiling identified fifteen differentially abundant proteins in normal and PSE-like broiler breast samples. Actin alpha, myosin heavy chain, phosphoglycerate kinase, creatine kinase M type, beta-enolase, carbonic anhydrase 2, proteasome subunit alpha, pyruvate kinase, and malate dehydrogenase were over-abundant (P < 0.05) in PSE-like broiler breast whereas phosphoglycerate mutase-1, alpha-enolase, ATP-dependent 6-phosphofructokinase, and fructose 1,6-bisphosphatase were over-abundant (P < 0.05) in normal meat. Thus, results indicated that differences in proteome abundance could be related to the meat quality differences between normal and PSE-like broiler breast meat.
Abstract
Background
Fatigue is one of the most important symptoms among patients receiving dialysis and is nominated as a core outcome to be reported in all clinical trials in this setting. However, ...few trials of interventions targeting fatigue have been conducted. Patients historically have rarely been involved in the design of interventions, which can limit acceptability and uptake. When asked, they have indicated a preference for lifestyle interventions, such as exercise, to improve fatigue. While some research has focussed on intradialytic exercise for patients receiving haemodialysis, patients have also indicated a preference for a convenient method of exercising with guidance, but on their own time outside of dialysis hours. In response to this, a mobile phone application was proposed as the method of delivery for a home-based exercise intervention targeting fatigue.
Methods
We convened a workshop with five breakout group sessions in Australia, with 24 patients on dialysis (16 haemodialysis and 8 peritoneal dialysis) and 8 caregivers to identify, prioritize and discuss exercise interventions for fatigue in patients receiving dialysis and the delivery of this through a mobile application.
Results
Of the 21 types of exercise identified, the top-ranked were walking outdoors, walking on a treadmill and cardio and resistance training. Six themes were identified: (i) ‘an expectation of tangible gains from exercise’, including strengthening and protecting against bodily deterioration related to dialysis; (ii) ‘overcoming physical limitations’, meaning that comorbidities, baseline fatigue and fluctuating health needed to be addressed to engage in exercise; (iii) ‘fear of risks’, which reinforced the importance of safety and compatibility of exercise with dialysis; (iv) ‘realistic and achievable’ exercise, which would ensure initial readiness for uptake; (v) ‘enhancing motivation and interest’ , which expected to support sustained use of the exercise intervention and (vi) ‘ensuring usability of the mobile application’ , which would require simplicity, convenience and comprehensibility.
Conclusion
Exercise interventions that are expected by patients to improve health outcomes and that are safe, realistic and easy to adopt may be more acceptable to patients on dialysis.
Dietary ractopamine improves pork leanness, whereas its effect on sarcoplasmic proteome has not been characterized. Therefore, the influence of ractopamine on sarcoplasmic proteome of post-mortem ...pork Longissimus thoracis muscle was examined. Longissimus thoracis samples were collected from carcasses (24h post-mortem) of purebred Berkshire barrows (n=9) managed in mixed-sex pens and fed finishing diets containing ractopamine (RAC; 7.4mg/kg for 14days followed by 10.0mg/kg for 14days) or without ractopamine for 28days (CON). Sarcoplasmic proteome was analyzed using two-dimensional electrophoresis and mass spectrometry. Nine protein spots were differentially abundant between RAC and CON groups. Glyceraldehyde-3-phosphate dehydrogenase and phosphoglucomutase-1 were over-abundant in CON, whereas serum albumin, carbonic anhydrase 3, l-lactate dehydrogenase A chain, fructose-bisphosphate aldolase A, and myosin light chain 1/3 were over-abundant in RAC. These results suggest that ractopamine influences the abundance of enzymes involved in glycolytic metabolism, and the differential abundance of glycolytic enzymes could potentially influence the conversion of muscle to meat.
•The effects of dietary ractopamine on pork sarcoplasmic proteome were examined.•Ractopamine influenced sarcoplasmic proteome profile of Longissimus thoracis.•The abundance of glycolytic enzymes was influenced by dietary ractopamine.
The effects of 4-hydroxy-2-nonenal (HNE) on redox stability of Oxy- and Deoxy- wild-type (WT) and recombinant sperm whale myoglobins (P88H/Q152H, L29F, H97A, and H64F) and hemin loss from ...Met-myoglobin (Mb) were investigated. HNE induced greater redox instability in WT and mutant Mbs compared to controls (p < 0.05). The extent of HNE-induced OxyMb oxidation was lesser in L29F (p < 0.05) and greater in H97A and P88H/Q152H than in WT (p < 0.05). H64F DeoxyMb was more redox stable than WT DeoxyMb in the presence of HNE (p < 0.05). HNE alkylation occurred exclusively on histidine residues, and histidine 48 was alkylated in all sperm whale myoglobins. HNE alkylation accelerated the protoporphyrin moiety loss only in H97A. Met- forms of WT and L29F but not Deoxy- or Oxy- forms released hemin during storage. Primary structure strongly influenced Mb redox stability in the presence of reactive secondary lipid oxidation products.
Previously, we showed that Abl family tyrosine kinases are activated by growth factors, and Abl is required for transition from G1 to S phase during PDGF-mediated proliferation. Here, we show that ...the SHP-2 tyrosine phosphatase, which acts to promote proliferation in response to cytokines and growth factors, is a novel substrate of endogenous Abl kinases during growth factor-mediated cellular proliferation. Using a pharmacological inhibitor and RNAi, we show that endogenous Abl kinases phosphorylate SHP-2 on Y580, and induce sustained activation of ERK kinases in response to growth factor stimulation in fibroblasts. Consistent with these data, SHP-2 is required for Abl-dependent PDGF-mediated proliferation since expression of an activated form of SHP-2 rescues the ability of Abl-Arg null fibroblasts to transit from G1 to S phase, whereas inhibition of SHP-2 signaling reduces the ability of Abl kinases to rescue the proliferation defect. Abl kinases also indirectly mediate phosphorylation of SHP-2 on Y63 and Y279, which are frequent sites of germline mutation in two cancer susceptibility syndromes. Significantly, we demonstrate that phosphorylation of SHP-2 on Y279 downregulates growth factor-induced sustained ERK activation and proliferation, supporting a role for Abl kinases not only in potentiating growth factor-mediated SHP-2 signaling, but also in negative-feedback regulation.
: Our overall objective was to better understand the effects of added pyruvate on enhanced beef color stability. The 2 possible mechanisms assessed were the role of pyruvate in lipid oxidation and ...direct interaction between pyruvate and beef myoglobin. Microsomes were incubated with pyruvate at pH 5.6, 25 °C, and lipid oxidation was measured hourly for 3 h. Bovine oxymyoglobin at pH 5.6 was incubated with pyruvate and used to quantify both redox stability (metmyoglobin formation) and pyruvate‐myoglobin adduction using mass spectrometry analysis. Surface color and lipid oxidation were measured on ground beef patties stored for 6 d in polyvinyl chloride over‐wrap (PVC) or high oxygen. Addition of pyruvate to microsomes decreased lipid oxidation compared with controls (P < 0.05). Conversely, no effect on myoglobin was observed (no changes in redox stability and no peaks corresponding to pyruvate were observed; P > 0.05). However, pyruvate increased color stability and decreased lipid oxidation of ground beef patties packaged in PVC and high oxygen. Pyruvate decreased nitric oxide metmyoglobin‐reducing capacity and oxygen consumption of patties compared with controls (P < 0.05). This research suggests that pyruvate may improve beef color stability primarily through its antioxidant effect on lipids.
Practical Application: Discoloration of meat often results in significant revenue loss. This study suggests that pyruvate can improve the color stability of patties packaged in high oxygen and PVC primarily through its antioxidant effect on lipids.
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