Context. Asteroseismology is a powerful tool to access the internal structure of stars. Apart from the important impact of theoretical developments, progress in this field has been commonly associated with the analysis of time-resolved observations. Recently, the so-called macroturbulent broadening has been proposed as a complementary and less expensive way - in terms of observational time - to investigate pulsations in massive stars. Aims: We assess to what extent this ubiquitous non-rotational broadening component which shapes the line profiles of O stars and B supergiants is a spectroscopic signature of pulsation modes driven by a heat mechanism. Methods: We compute stellar main-sequence and post-main-sequence models from 3 to 70 M⊙ with the ATON stellar evolution code, and determine the instability domains for heat-driven modes for degrees ℓ = 1-20 using the adiabatic and non-adiabatic codes LOSC and MAD. We use the observational material compiled in the framework of the IACOB project to investigate possible correlations between the single snapshot line-broadening properties of a sample of ≈260 O and B-type stars and their location inside or outside the various predicted instability domains. Results: We present an homogeneous prediction for the non-radial instability domains of massive stars for degree ℓ up to 20. We provide a global picture of what to expect from an observational point of view in terms of the frequency range of excited modes, and we investigate the behavior of the instabilities with respect to stellar evolution and the degree of the mode. Furthermore, our pulsational stability analysis, once compared to the empirical results, indicates that stellar oscillations originated by a heat mechanism cannot explain alone the occurrence of the large non-rotational line-broadening component commonly detected in the O star and B supergiant domain. Based on observations made with the Nordic Optical Telescope, operated by NOTSA, and the Mercator Telescope, operated by the Flemish Community, both at the Observatorio del Roque de los Muchachos (La Palma, Spain) of the Instituto de Astrofísica de Canarias.