Ti6Al4V alloy was shot peened by using stainless-steel shots with different sizes (0.09–0.14 mm (S10) and 0.7–1.0 mm (S60)) for two durations (5 and 15 min) using a custom-designed peening system. The shot size was the main parameter modifying the roughness (0.74 µm for S10 vs. 2.27 µm for S60), whereas a higher peening time slightly increased roughness. Hardness improved up to approximately 35% by peening with large shots, while peening time was insignificant in hardness improvement. However, longer peening duration with large shots led to an unwanted formation of micro-cracks and delamination on the peened surfaces. After dry sliding wear tests, the mass loss of peened samples (S60 for 15 min) was 25% higher than that of un-peened samples, while the coefficient of friction decreased by 12%. Plastically deformed regions and micro-scratches were observed on the worn surfaces, which corresponds to mostly adhesive and abrasive wear mechanisms. The present study sheds light on how surface, subsurface and tribological properties of Ti6Al4V vary with shot peening and peening parameters, which paves the way for the understanding of the mechanical, surface, and tribological behavior of shot peened Ti6Al4V used in both aerospace and biomedical applications.