Heat stress is a serious and widespread threat to the quality and yield of many crops, including grape (Vitis vinifera), which is cultivated worldwide. Here, we conducted phosphoproteomic and acetylproteomic analyses of leaves from grape cultivated in four distinct temperature regimes. The phosphorylation or acetylation of a total of 1,011 phosphoproteins with 1,828 phosphosites, and 96 acetylproteins with 148 acetylsites, changed when plants were grown at 35°C, 40°C and 45°C in comparison with the proteome profiles of plants grown at 25°C. The greatest number of changes were observed at higher temperatures. Functional classification and enrichment analysis indicated that phosphorylation, rather than acetylation, of serine/arginine-rich splicing factors was involved in the responses to high temperatures. This is congruent with previous observations that alternative splicing events occurred more frequently in grapevine under high temperature. Changes in acetylation patterns were more common than changes in phosphorylation patterns in photosynthesis-related proteins at high temperatures, while heat shock proteins were more associated with modifications in phosphorylation than in acetylation. Nineteen proteins were identified with changes in both phosphorylation and acetylation, consistent with crosstalk between these post-translational modifications.