Proteomic Analysis of the Effect of Proteasome on Quality Changes in Qinchuan Beef during Postmortem Storage

In this study, the Longissimus dorsi muscle of Qinchuan beef stored at 4 ℃ after slaughter was determined for its quality indicators, energy level and proteasome activity. Meanwhile, differentially expressed proteins (DEPs) associated with proteasomes were identified. Bioinformatics methods were used to analyze the response mechanism of proteasomes to the quality and energy metabolism of postmortem beef. The results showed that pH decreased and then increased with increasing storage time, b* and myofibrillar fragmentation index (MFI) increased, L^*, a^*, centrifugal loss, cooking loss and shear force increased first and then decreased, the contents of the basic energy substances ATP, ADP and AMP decreased, and the relative activity of the 20S proteasome decreased significantly (P < 0.05). The correlation analysis showed that the activity of the 20S proteasome was significantly negatively correlated with b* (P < 0.05), positively correlated with ADP content (P < 0.05), positively correlated with AMP content (P < 0.01), and negatively correlated with MFI (P < 0.01), indicating a close relationship between proteasome activity and energy metabolism as well as beef quality. Using 4D label-free quantitative (LFQ) proteomics, eight differentially expressed proteasome subunits and 27 DEPs were identified. Gene Ontology (GO) annotation analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed that these proteasome subunits and related DEPs had molecular functions such as endopeptidase activity, actin binding, microfilament motor activity, and were involved in biological processes such as ubiquitin-dependent proteolytic metabolism, skeletal muscle contraction, muscle contraction, and glycolysis, thus causing changes in the quality of Qinchuan beef. During the early postmortem period, the proteasomes regulated the biological pathways by consuming energy substances to catabolize proteins, and during the later period, when the energy substances were depleted, the proteasomes catabolized proteins in postmortem beef through the energy provided by glycolysis and non-ATP-dependent proteasome subunits, which eventually affected the quality of Qinchuan beef.