scholarly journals Tandem Mass Tag Labeling to Identify Proteome Changes in Beef Longissimus Lumborum and Psoas Major Muscles During Early Postmortem Period

2019 ◽  
Vol 3 (2) ◽  
Author(s):  
C. Zhai ◽  
B. A. Djimsa ◽  
J. E. Prenni ◽  
R. J. Delmore ◽  
D. R. Woerner ◽  
...  
Keyword(s):  
Meat Quality ◽  
Color Stability ◽  
Quality Attributes ◽  
Tandem Mass ◽  
Tandem Mass Tag ◽  
Proteome Profile ◽  
Apoptotic Proteins ◽  
Psoas Major ◽  
Differentially Abundant Proteins ◽  
Early Postmortem

ObjectivesLongissimus lumborum (LL) and psoas major (PM) are important muscles in beef hindquarters that exhibit variation in meat quality attributes. Postmortem metabolism (muscle-to-meat conversion) affects biochemical properties of muscles and in turn influence the meat quality. Although previous research has indicated that variation in the proteome profile of LL and PM post-rigor influences meat quality attributes such as tenderness and color stability during retail display, limited research has examined the influence of early postmortem metabolism on meat quality. Tandem mass tag (TMT) labeling is a chemical labeling approach used for accurate mass spectrometry-based quantification and identification of biological macromolecules. Therefore, the objective of this study was to use TMT labeling to examine proteome profile variation between beef LL and PM during the early postmortem period.Materials and MethodsMuscle biopsy samples were collected from carcasses (n = 4) at 45 min, 12 h, and 36 h postmortem from a commercial beef processing facility. Samples were frozen immediately in liquid nitrogen and stored at –80°C until proteomic analysis. Proteome was analyzed using TMT label containing ten different isobaric compounds with the same mass and chemical structure composed of an amine-reactive NHS-ester group, a spacer arm, and a mass reporter. After labeling and peptide fractionation, all the samples were multiplexed and ran through the Orbitrap Velos mass spectrometer equipped with a Nanospray Flex ion source to identify differentially abundant proteins. The proteins exhibiting 1.5-fold or more intensity difference and a statistical difference (P < 0.05) between LL and PM or within the muscles during the postmortem were reported as differentially abundant.ResultsSeventy differentially abundant proteins (P < 0.05) were identified from three comparisons between the muscles (31 proteins in PM 45 min vs. LL 45 min, 41 proteins in PM 12 h vs. LL 12 h, 49 proteins in PM 36 h vs. LL 36 h). However, no difference (P > 0.05) in protein expression within a muscle was observed during these time points. The differentially abundant proteins were mainly involved in oxidative phosphorylation and ATP-related transport, tricarboxylic acid cycle, NADPH regeneration, fatty acid degradation, muscle contraction, calcium signaling, chaperone activity, oxygen transport, as well as degradation of the extracellular matrix. At early postmortem, overabundant anti-apoptotic proteins in LL could cause high metabolic stability, enhanced autophagy, and delayed apoptosis, while overabundant metabolic enzymes and pro-apoptotic proteins in PM could accelerate the reactive oxygen species generation and programmed cell death.ConclusionDifferentially abundant proteins between LL and PM during the early postmortem were primarily associated with cellular metabolism and programmed cell death. The greater oxidative and color stability in LL compared to PM could be related to the increased expression of anti-apoptotic proteins and the decreased expression of metabolic enzymes and proapoptotic proteins in LL.

scholarly journals Tandem Mass Tag Labeling to Identify Proteome Changes in Beef Longissimus Lumborum and Psoas Major Muscles During Early Postmortem Period

2019 ◽  
Vol 3 (2) ◽  
pp. 167-167
Author(s):  
C. Zhai ◽  
B. A. Djimsa ◽  
J. E. Prenni ◽  
R. J. Delmore ◽  
D. R. Woerner ◽  
...  
Keyword(s):  
Tandem Mass ◽  
Tandem Mass Tag ◽  
Longissimus Lumborum ◽  
Proteome Changes ◽  
Psoas Major ◽  
Early Postmortem ◽  
Mass Tag

scholarly journals Tandem Mass Tag Labeling-Based Analysis to Characterize Muscle-Specific Proteome Changes during Postmortem Aging of Bison Longissimus Lumborum and Psoas Major Muscles

2021 ◽  
Author(s):  
MD Mahmudul Hasan ◽  
Mahamud-ur Rashid ◽  
Surendranath P Suman ◽  
Helene Perreault ◽  
Jitendra Paliwal ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Fatty Acid ◽  
Oxygen Transport ◽  
Color Stability ◽  
Acid Oxidation ◽  
Tandem Mass ◽  
Tandem Mass Tag ◽  
Postmortem Aging ◽  
Longissimus Lumborum ◽  
Psoas Major

The objective of the study was to examine the variations in sarcoplasmic proteomes of bison longissimus lumborum (LL) and psoas major (PM) muscles during postmortem aging utilizing tandem mass tag (TMT) isobaric labeling coupled with liquid chromatography mass-spectrometry (LC-MS/MS) for the categorization of muscles with muscle-specific inherent color stability. A total of 576 proteins were identified (P < 0.05) in both bison LL and PM muscles, where 97 proteins were identified as differentially abundant (fold change > 1.5, P < 0.05) from the three comparisons between muscles during postmortem aging periods (PM vs LL at 2 d, 7 d and 14 d). Among those proteins, the most important protein groups based on functions are related to electron transport chain (ETC) or oxidative phosphorylation, tricarboxylic acid cycle (TCA), ATP transport, carbohydrate metabolism, fatty acid oxidation, chaperones, oxygen transport, muscle contraction, calcium signaling, and protein synthesis. In PM, most of the proteins from ETC, TCA cycle, fatty acid oxidation, ATP and oxygen transport, and muscle contraction were more abundant or exhibited increased expression during aging compared to LL. On the other hand, the proteins involved in carbohydrate metabolism, chaperone function and protein synthesis mostly exhibited decreased expression in PM muscle relative to LL. These results clearly demonstrate that the proteins associated with oxidative metabolism showed increased expression in PM muscles. This indicates that oxidative damage or subsequent color deterioration resulted in bison PM muscles being attacked by the reactive oxygen species produced during those metabolic process. In contrast, proteins involved in glycolysis and chaperone activity exhibited a decrease in expression in bison PM muscles, resulting decline in color stability compared with LL. Because glycolytic enzymes and chaperones maintain oxidative and/or color stability by producing reducing equivalents in glycolytic pathway and with the protein folding ability of chaperones, respectively in LL muscles.

Quantitative phosphoproteomic analysis among muscles of different color stability using tandem mass tag labeling

2018 ◽  
Vol 249 ◽  
pp. 8-15 ◽  
Author(s):  
Zheng Li ◽  
Meng Li ◽  
Xin Li ◽  
Jianzeng Xin ◽  
Ying Wang ◽  
...  
Keyword(s):  
Color Stability ◽  
Tandem Mass ◽  
Tandem Mass Tag ◽  
Different Color ◽  
Mass Tag

scholarly journals Elucidating the involvement of apoptosis in postmortem proteolysis in porcine muscles from two production cycles using metabolomics approach

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Danyi Ma ◽  
Dong Ho Suh ◽  
Jiaying Zhang ◽  
Yufan Chao ◽  
Alan W. Duttlinger ◽  
...  
Keyword(s):  
Meat Quality ◽  
Troponin T ◽  
Antioxidant Metabolites ◽  
Production Cycles ◽  
Branched Chain ◽  
Underlying Mechanisms ◽  
Psoas Major ◽  
Quality Variation ◽  
Early Postmortem

AbstractApoptosis has been suggested as the first step in the process of conversion of muscle into meat. While a potential role of apoptosis in postmortem proteolysis has been proposed, the underlying mechanisms by which metabolome changes in muscles would influence apoptotic and proteolytic process, leading to meat quality variation, has not been determined. Here, apoptotic and proteolytic attributes and metabolomics profiling of longissimus dorsi (LD) and psoas major (PM) muscles in pigs from two different production cycles (July–Jan vs. Apr–Sep) were evaluated. PM showed higher mitochondrial membrane permeability (MMP), concurrent with less extent of calpain-1 autolysis and troponin T degradation and higher abundance of HSP27 and αβ-crystallin compared to LD (P < 0.05). Apr–Sep muscles showed concurrence of extended apoptosis (indicated by higher MMP), calpain-1 autolysis and troponin T degradation, regardless of muscle effects (P < 0.05). Metabolomics profiling showed Apr–Sep muscles to increase in oxidative stress-related macronutrients, including 6-carbon sugars, some branched-chain AA, and free fatty acids. Antioxidant AA (His and Asp) and ascorbic acid were higher in July–Jan (P < 0.05). The results of the present study suggest that early postmortem apoptosis might be positively associated with pro-oxidant macronutrients and negatively associated with antioxidant metabolites, consequently affecting meat quality attributes in a muscle-specific manner.

Tandem mass tag labeling to characterize muscle-specific proteome changes in beef during early postmortem period

2020 ◽  
Vol 222 ◽  
pp. 103794 ◽  
Author(s):  
Chaoyu Zhai ◽  
Blanchefort A. Djimsa ◽  
Jessica E. Prenni ◽  
Dale R. Woerner ◽  
Keith E. Belk ◽  
...  
Keyword(s):  
Tandem Mass ◽  
Tandem Mass Tag ◽  
Proteome Changes ◽  
Early Postmortem ◽  
Mass Tag

scholarly journals Endpoint Temperature Influences Sarcoplasmic Proteome Profile of Cooked Beef Longissimus Lumborum

2019 ◽  
Vol 3 (2) ◽  
Author(s):  
A. P. A. A. Salim ◽  
S. P. Suman ◽  
S. Li ◽  
Y. Wang ◽  
J. Chen ◽  
...  
Keyword(s):  
Color Stability ◽  
Fructose Bisphosphate ◽  
Cooking Temperature ◽  
Longissimus Lumborum ◽  
Fructose Bisphosphate Aldolase ◽  
C 71 ◽  
Proteome Profile ◽  
Aldolase B ◽  
Differentially Abundant Proteins ◽  
Cooked Color

ObjectivesCooking ensures safety and enhances the palatability attributes of meat. Denaturation of myoglobin results in the dull-brown color of cooked meats. The denaturation of sarcoplasmic proteins is influenced by the degree of heat treatment, and their solubility is decreased with an increase in the endpoint cooking temperature. While previous studies examined the relationship between myoglobin denaturation, cooked color, and internal temperature in beef, investigations are yet to be undertaken to characterize the association between endpoint temperature, sarcoplasmic proteome, and color attributes in cooked steaks. Therefore, the objective of the present study was to examine the influence of endpoint cooking temperature (60 and 71°C) on sarcoplasmic proteome and internal color of beef longissimus lumborum (LL) steaks.Materials and MethodsEight (n = 8) beef LL muscles (14 d postmortem; USDA Choice) were obtained from a commercial packing plant. Two 2.5-cm thick steaks were fabricated from the center of the muscles and were cooked to internal endpoint temperature of 60°C (C-60) or 71°C (C-71) in a clam-shell grill. Cooked steaks were immediately cooled in slushed ice, sliced parallel to the grilled surface, and internal redness (a* value) and color stability (R630/580) were evaluated instrumentally. Sarcoplasmic proteome from the interiors of the cooked steaks was analyzed using 2-dimensional electrophoresis, and the gel images were digitally analyzed. The protein spots exhibiting more than 2.5-fold intensity differences (P < 0.05) between C-60 and C-71 were subjected to in-gel tryptic digestion and were identified by tandem mass spectrometry.ResultsThe C-60 steaks demonstrated greater (P < 0.05) a* and R630/580 than their C-71 counterparts. Seven differentially abundant proteins were identified and were over-abundant (P < 0.05) in C-60 compared to C-71. The differentially abundant proteins belong to 6 functional groups, i.e., transport proteins (serum albumin and hemoglobin), energy metabolism (adenylate kinase isoenzyme 1), chaperones (heat shock protein β-1), antioxidant (thioredoxin-dependent peroxide reductase), glycolytic enzymes (fructose-bisphosphate aldolase B), and protease (cytosol aminopeptidase).ConclusionThe findings indicated that the endpoint cooking temperature influences the internal cooked color and the sarcoplasmic proteome profile of beef LL steaks. The overabundant proteins in steaks cooked to 60°C may be utilized as potential biomarkers for undercooked beef, which is a source for foodborne infections.

scholarly journals Effect of Different Photoperiods on Quality Attributes and Oxidative Stability of Breast Meat (M. Pectoralis Major) from Broilers

2019 ◽  
Vol 3 (2) ◽  
Author(s):  
E. Will ◽  
J. R. Tuell ◽  
J. Park ◽  
W. Wang ◽  
H.-W. Cheng ◽  
...  
Keyword(s):  
Oxidative Stability ◽  
Meat Quality ◽  
Color Stability ◽  
Quality Attributes ◽  
Water Holding Capacity ◽  
Moisture Loss ◽  
Breast Meat ◽  
Significant Difference ◽  
Thiol Content ◽  
Water Holding

ObjectivesIn the broiler industry, high photoperiod (the duration of light exposure per day) regimes have traditionally been utilized to increase yield of breast meat, as well as maximize feed intake and growth rate in the growing period. However, recent literature reports this practice may have adverse effects on broiler welfare, resulting in impaired mobility and increased incidence of leg abnormalities. However, little information available in the current literature regarding effects of photoperiod on meat quality attributes and oxidative stability of broiler meat. Thus, the objective of this study was to evaluate the quality characteristics and lipid/protein oxidative stability of breast meat from broilers that were exposed to different photoperiod combinations.Materials and MethodsA total of 432 Ross 308 broiler chicks were allocated among 4 rooms each subjected to one of the following photoperiod treatments (hours Light: Dark): 20L:4D, 18L:6D, 16L:8D and 12L:12D, with 6 pens per treatment. At d 42, 2 broilers per pen (12 broilers/treatment) were randomly selected, harvested and air chilled for 24 h at 2°C. At l day postmortem, paired breast muscles (M. pectoralis) were collected for the meat quality analyses such as, water-holding capacity (drip/purge/cook loss), Warner-Bratzler shear force (WBSF), and display color for 7 d under fluorescent light (1450 lx). Lipid oxidation was assessed via the TBARS assay and protein oxidation by thiol content measured at d 1 and d 7 of display. The experimental design was randomized complete block design. Data were analyzed using the PROC MIXED procedure of SAS, and means were separated using least significant differences (P < 0.05).ResultsPhotoperiod had no effect on fillet yield and pH (P > 0.05). No significant difference in WBSF was found between treatments, although 12L:12D had a trend of higher WBSF (P = 0.08). Higher moisture loss during carcass chilling was found in carcasses from 20L:4D compared to 16L:8D and 12L:12D treatments (P < 0.05). No other measure of water-holding capacity was affected, though the 16L:8D treatment demonstrated a trend of higher freezing/thawing loss compared to other treatments (P = 0.06). Proximate moisture, protein and lipid contents were unaffected by photoperiod (P > 0.05), but higher ash was observed in 16L:8D over 20L:4D and 18L:6D (P < 0.05). Different photoperiod combinations affected color stability of breast meat during display (P < 0.05). Of note, fillets from 20L:4D maintained highest L* and hue angle, and least a* values (P < 0.05), indicating inferior color stability compared to other treatments. Oxidation increased with display, and fillets from 20L:4D and 18L:6D had higher TBARS over 12L:12D (P < 0.05); no photoperiod effect was observed in thiol content (P > 0.05).ConclusionResults suggest 20L:4D photoperiod regimes may be detrimental to meat quality, as carcasses from this treatment group had higher moisture loss during chilling, and color measurements characterized these fillets as being paler and more discolored than other treatments. Fillets from 12L:12D maintained lower TBARS than 20L:4D and 18L:6D, suggesting photoperiod regimes allowing more hours of dark may be beneficial in improving oxidative stability. Further studies determining the effects of photoperiod on quality and protein functionality of chicken meat for processing technological would be highly warranted.

TMT-Based Quantitative Proteomic Analysis Identified Proteins and Signaling Pathways Involved in the Response to Xanthatin Treatment in Human HT-29 Colon Cancer Cells

2021 ◽  
Vol 21 ◽  
Author(s):  
Yadi Geng ◽  
Lingli Li ◽  
Ping Liu ◽  
Zhaolin Chen ◽  
Aizong Shen ◽  
...  
Keyword(s):  
Colon Cancer ◽  
Cancer Cells ◽  
Tandem Mass ◽  
Colon Cancer Cells ◽  
Comprehensive Understanding ◽  
Antitumor Effects ◽  
Tandem Mass Tag ◽  
Ht 29 ◽  
Differentially Abundant Proteins ◽  
Mass Tag

Background: Xanthatin is a plant-derived bioactive sesquiterpene lactone from the Xanthium strumarium L., and it has been used as a traditional Chinese medicine. Recently, many studies have reported that xanthatin has anticancer activity. However, a comprehensive understanding of the mechanism underlying the antitumor effects of xanthatin is still lacking. Objective: To systematically and comprehensively identify the underlying mechanisms of xanthatin on cancer cells, quantitative proteomic techniques were performed. Methods: Xanthatin induced HT-29 colon cancer cells death was detected by lactate dehydrogenase (LDH) release cell death assay. Differentially abundant proteins in two groups (control groups and xanthatin treatment groups) of human HT-29 colon cancer cells were identified using tandem mass tag (TMT) quantitative proteomic techniques. All the significant differentially abundant proteins were generally characterized by performing hierarchical clustering, Gene Ontology (GO) enrichment analyses and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. We chose Western blot analysis to validate the candidate proteins in the proteomics results. Results: A total of 5637 proteins were identified, of which 397 significantly differentially abundant proteins in the groups were quantified. Based on the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses, we found that p53-related signaling played an important role in xanthatin-treated HT-29 colon cancer cells. p53-upregulated modulator of apoptosis (Puma), Sestrin-2 and p14ARF, which were selected from among p53-related signaling proteins, were further validated, and the results were consistent with the tandem mass tag quantitative proteomic results. Conclusion: We first investigated the molecular mechanism underlying the effects of xanthatin treatment on HT-29 colon cancer cells using tandem mass tag quantitative proteomic methods and provided a global comprehensive understanding of the antitumor effects of xanthatin. However, it is necessary to further confirm the function of the differentially abundant proteins and the potentially associated signaling pathways.

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