This study aimed to evaluate the effect of different aging times (1, 6 and 14 days) on the tenderization rate and protein changes in ten Martina Franca donkey striploins using a proteomic approach. During aging, a progressive decrease in share force, hardness, gumminess, and chewiness together with an increase in myofibril fragmentation index were observed. Proteolysis monitored by immunoblotting revealed a progressive degradation of desmin and fast troponin-T over time and an increase of their degradation products up to 14 days aging. Proteomics revealed by means of two-dimensional electrophoresis 37 protein spots corresponding to 15 proteins to change significantly by increasing aging time. These proteins belong to three pathways, these being “muscle contraction, structure, and associated proteins” (9 proteins); ii) “energy metabolism” (5 proteins); and iii) “chaperone” (1 protein). This study is the first to highlight the possible role of interconnected pathways in driving the final quality of donkey meat and predicting its texture.