Effect of oxidation, pH, and ionic strength on calpastatin inhibition of μ- and m-calpain

Carlin, K.R. Maddock; Huff-Lonergan, Elisabeth J.; Rowe, L. J.; Lonergan, Steven M.

doi:10.2527/2006.844925x

Cited by 85 publications

(61 citation statements)

References 39 publications

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“…The rate of activation and autolysis of calpain-1 could impact the amount of protein degradation by calpain-1. Calpain-1 activity is inhibited as pH declines from pH 7 to pH 6 and below (Carlin et al, 2006;Bee et al, 2007) or by oxidative conditions (Rowe et al, 2004a;Chen et al, 2014). Postmortem pH decline data are not available for these loins, but if differences in pH decline actually existed, they could potentially explain the differences in proteolysis exhibited between classification groups in this study.…”

Section: Discussionmentioning

confidence: 74%

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Postmortem protein degradation is a key contributor to fresh pork loin tenderness

Carlson¹,

Prusa²,

Fedler³

et al. 2017

Journal of Animal Science

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The objective of this study was to determine factors that influence tenderness independent of variation in pH, color, or marbling. To achieve the objective, 2 sample groups were chosen from a population of 159 pork loins aged 11 to 16 d. Predetermined ranges (ultimate pH, 5.54 to 5.86; marbling score, 1.0 to 3.0; percent total lipid, 1.61 to 3.37%) were defined for inclusion of individual loins in the study. The pork loins with the greatest (n = 12) and least (n = 12) Instron star probe values were assigned to 2 classification groups. The high star probe group had an average star probe that was 2.8 kg greater than the low star probe group (7.75 vs. 4.95 kg). Pork quality and sensory characteristics of pH, subjective and instrumental color values, cook loss, sensory tenderness, chewiness, juiciness, pork flavor, and off flavor were determined on fresh, never frozen pork chops. Lipid content, sarcomere length, myosin heavy-chain profile, and calpain autolysis were determined. Degradation of troponin-T, desmin, filamin, and titin were evaluated on the protein extracts from each sample. Pork loin pH, subjective color scores, Minolta L values, sarcomere length, and myosin heavychain composition were not different across groups. Chops from the low star probe group had a significantly greater marbling score (2.3 vs. 1.9) and lipid content (2.61 vs. 2.23%). Calpain-1 was completely autolyzed in both high and low star probe samples, demonstrating that calpain-1 potentially had been active in all samples. Low star probe whole-muscle protein extracts had more troponin-T (P < 0.01), desmin (P < 0.01), and filamin degradation (P < 0.01) than high star probe samples. Both classification groups showed degradation of titin. Remarkably, some high star probe samples still had observable intact bands of titin on SDS-PAGE gels. These results demonstrate that significant variation in instrumental tenderness is observed within a moderate pH range. Lipid content and proteolysis both appear to contribute to this variation.

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Section: Discussionmentioning

confidence: 74%

“…Oxidation of desmin changes the secondary structure of desmin, which increases susceptibility to proteolysis by caspases (Chen et al, 2014). Oxidation is also known to decrease the activity of calpain-1, ultimately reducing desmin degradation by calpain-1 (Rowe et al, 2004b;Carlin et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

Postmortem protein degradation is a key contributor to fresh pork loin tenderness

Carlson¹,

Prusa²,

Fedler³

et al. 2017

Journal of Animal Science

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“…Oxidation of calpain can create a disulfide bond at the active site of the protease, resulting in inhibition of its activity, which is reversible in the presence of reducing conditions (Lametsch et al, 2008). However, oxidation also appears to decrease the efficacy of the inhibitor of calpain, calpastatin, at physiological pH (Carlin et al, 2006). Damaged proteins are also conveyed directly to the proteasome by molecular chaperones that include heat shock proteins such as heat shock protein 90 and α-crystallin B chain (Glickman and Ciechanover, 2002).…”

Section: Methodsmentioning

confidence: 99%

Evidence of decreased muscle protein turnover in gilts selected for low residual feed intake1

Cruzen

Harris

Hollinger

et al. 2013

Journal of Animal Science

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The objective of this study was to evaluate the contribution of muscle protein turnover (synthesis and degradation) to the biological basis for genetic differences in finisher pigs selected for residual feed intake (RFI). Residual feed intake is defined as the difference between expected feed intake (based on the achieved rate of BW gain and backfat depth of individual pigs) and the observed feed intake of the individual pig. We hypothesized that protein turnover would be reduced in pigs selected for low RFI. Twelve gilts from a line selected for 7 generations for low RFI and 12 from a contemporary line selected for 2 generations for high RFI were paired by age and BW and fed a standard corn-soybean diet for 6 wk. Pigs were euthanized, muscle and liver samples were collected, and insulin signaling, protein synthesis, and protein degradation proteins were analyzed for expression and activities. Muscle from low RFI pigs tended to have less μ-and m-calpain activities (P = 0.10 and 0.09, respectively) and had significantly greater calpastatin activity and a decreased μ-calpain:calpastatin activity ratio (P < 0.05). Muscle from low RFI pigs had less 20S proteasome activity compared with their high RFI counterparts (P< 0.05). No differences in insulin signaling intermediates and translation initiation signaling proteins [mammalian target of rapamycin (mTOR) pathway] were observed (P > 0.05). Postmortem proteolysis was determined in the LM from the eighth generation of the low RFI pigs versus their high RFI counterparts (n = 9 per line). Autolysis of μ-calpain was decreased in the low RFI pigs and less troponin-T degradation product was observed at 3 d postmortem (P < 0.05), indicating slowed postmortem proteolysis during aging in the low RFI pigs. These data provide significant evidence that less protein degradation occurs in pigs selected for reduced RFI, and this may account for a significant portion of the increased efficiency observed in these animals. KeywordsSwine Feed Efficiency, calpain, calpastatin, proteasome, protein synthesis, residual feed intake, swine ABSTRACT: The objective of this study was to evaluate the contribution of muscle protein turnover (synthesis and degradation) to the biological basis for genetic differences in finisher pigs selected for residual feed intake (RFI). Residual feed intake is defined as the difference between expected feed intake (based on the achieved rate of BW gain and backfat depth of individual pigs) and the observed feed intake of the individual pig. We hypothesized that protein turnover would be reduced in pigs selected for low RFI. Twelve gilts from a line selected for 7 generations for low RFI and 12 from a contemporary line selected for 2 generations for high RFI were paired by age and BW and fed a standard corn-soybean diet for 6 wk. Pigs were euthanized, muscle and liver samples were collected, and insulin signaling, protein synthesis, and protein degradation proteins were analyzed for expression and activities. Muscle from low RFI pigs tended to have less ...

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“…Ot can also cause the inactivation of proteolytic enzymes involved in the meat tenderization process. All of this leads to a decrease in the proteolytic susceptibility (Carlin et al, 2006). This explains the lower proteolytic activity (MFO) observed in PFN and Normal meats, which gave them a firmer texture in relation to PSE meat (Table 1).…”

Section: Characterization Of Broiler Breastmentioning

confidence: 89%

Further evidence for the existence of broiler chicken PFN (pale, firm, non-exudative) and PSE (pale, soft, exudative) meat in brazilian commercial flocks

et al. 2018

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This work aims to investigate the incidence of two color abnormalities in broiler meat: PFN (Pale, Firm, Non-exudative) and PSE (Pale, Soft, Exudative), as well as the physicochemical alterations promoted by those anomalies. The samples of broiler breast meat (n = 700) were classified as Normal (445.8), PSE (L*≥53, pH<5.8) and PFN (L*≥53; pH>5.8). The occurrence of Normal, PSE and PFN samples was 69%, 11% and 20%, respectively. PFN samples presented a 4.2% higher water-holding capacity compared to PSE meat. PSE meats displayed a higher MFO (89.4) when compared to PFN and Normal meats. Ot was also noticed that the levels of lipid oxidation were greater in PSE breasts compared to Normal breasts. The total carbonyl in PFN, Normal and PSE meats was 8.2, 7.4 and 5.7 nmol/mg proteins, respectively. These results confirm the existence of a PFN anomaly in broiler breast meat, which presents functional properties similar to the Normal group. On addition to that, the less firm texture displayed by PSE meats is the result of a greater proteolytic activity, which seems to be related not only to the activation of the calpains, due to the excessive Ca 2+, but also to the lower level of protein oxidation.Keywords: abnormal coloration; functional properties; lipid oxidation; protein oxidation. Practical Application: Data on incidence and characterization of abnormal color meats are important for the prevention and understanding of these anomalies.

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Effect of oxidation, pH, and ionic strength on calpastatin inhibition of μ- and m-calpain

Cited by 85 publications

References 39 publications

Postmortem protein degradation is a key contributor to fresh pork loin tenderness

Postmortem protein degradation is a key contributor to fresh pork loin tenderness

Evidence of decreased muscle protein turnover in gilts selected for low residual feed intake1

Further evidence for the existence of broiler chicken PFN (pale, firm, non-exudative) and PSE (pale, soft, exudative) meat in brazilian commercial flocks

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