This study used proteomic analysis to evaluate how four cooking methods impact the quality of beef from Rikaze humped cattle. Conventional boiling, high-pressure boiling, roasting and frying produced varying numbers of differentially abundant proteins (DAPs) in Longissimus dorsi and hind leg beef samples compared to the raw meat. Principal component analysis, hierarchical clustering and correlation analysis revealed 11 and 18 proteins significantly associated with meat tenderness and colour, respectively. Myosin heavy chain 7 (MYH7), myosin light chain 2 (MYL2) and myosin light chain 6B (MYL6B), involved in the sarcomere (GO:0030017) and cardiac muscle contraction (map04260) pathways were negatively correlated with tenderness, indicating that their decreased abundance contributes to improved tenderness after cooking. The cytoskeletal protein α-crystallin B chain (CRYAB) showed a positive correlation with shear force, suggesting a role in toughness. PGAM2, ALDOA and PKM, participating in ADP metabolism (GO:0046031) and glycolysis (map00010), influenced meat texture, while glycolytic enzymes ALDOA, CKM and PKM promoted changes in lightness (L*). MYBPC2 and VDAC2 were negatively correlated with L*. EEF1G, PGP, PPIA and PRDX2 were negatively correlated with redness (a*), and DES was positively correlated with yellowness (b*). Overall, wet-heat cooking altered mainly skeletal, heat-shock and cytoskeletal proteins and enhanced tenderness, while proteins related to energy metabolism and oxidative stress were closely linked to colour development. These key proteins could serve as potential biomarkers for predicting the eating quality of humped cattle beef and provide a basis for the development of high-value beef products.
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