The microstructure, mechanical properties, and deformation behavior of wrought and electron beam additive manufactured (EBAM) Ti-6Al-4V samples under scratching were studied. As-received wrought Ti-6Al-4V was subjected to thermal treatment to obtain the samples with microstructure and mechanical characteristics similar to those of the EBAM samples. As a result, both alloys consisted of colonies of α phase laths within prior β phase grains and were characterized by close values of hardness. At the same time, the Young’s modulus of the EBAM samples determined by nanoindentation was lower compared with the wrought samples. It was found that despite the same hardness, the scratch depth of the EBAM samples under loading was substantially smaller than that of the wrought alloy. A mechanism was proposed, which associated the smaller scratch depth of EBAM Ti-6Al-4V with α′→α″ phase transformations that occurred in the contact area during scratching. Ab initio calculations of the atomic structure of V-doped Ti crystallites containing α or α″ phases of titanium were carried out to support the proposed mechanism.