Beef longissimus dorsi muscle with different high temperature treatment (110 ℃, 115 ℃ and 121 ℃, respectively, heating 3, 6, 9, 12 and 15 min) were tested for time and temperature-related changes in different solubility and structural proteins, total nitrogen and non-protein nitrogen, amino nitrogen content, proteolysis index and SDS-PAGE patterns of total muscle protein during high temperature processing. The results showed that the contents of water-soluble protein and salt-soluble protein decreased distinctly with the increase of heating temperature and time. After heating at 121 ℃ for 15 min, the content of sarcoplasmic protein in the beef longissimus dorsi muscle was decreased by 86.9%, myofibrillar protein decreased by 89.88%. The content of total nitrogen and amino nitrogen increased first and then decreased. The effect of heating time on non-protein nitrogen and protein hydrolysis index was significant (P＜0.05). The myosin heavy chain band of 200 kDa gradually disappeared as the heating time prolonged, while the actin band of 44.3 kDa did not change significantly during the high temperature treatment.