The purpose of this research was to investigate the causes and consequences of pHu variations in beef cattle. For that, were evaluated 176 Nellore beef cattle, and classified into two different pHu groups: High (≥6.0, N=17) and Normal (<5.8, N=159). Plasma concentrations of cortisol and adrenocorticotropic hormone, lactate and glycogen muscular content, meat color, shear force and Longissimus thoracis muscle proteomic profile were evaluated and compared between pHu groups. Muscle glycogen content, meat color and shear force statistically differed between pHu groups. Label-free quantitative proteomic analysis revealed ten differentially abundant proteins between pHu groups, involved in metabolic processes and muscle contraction. Thirty-six and 31 proteins were exclusively present in Normal and High pHu group, respectively, which were related to TCA cycle, cortisol production, calcium regulation, and antioxidant function. The MYH7, UGP2, and VDAC3 were identified as potential indicators of pHu variations. CALM and NNT appeared to be interesting proteins to better understand the metabolic pathways behind pHu.