In the last decade mass spectrometry-based methods emerged for the quantitative profiling of dynamic changes in protein phosphorylation, monitoring the behavior of thousands of phosphorylation sites in a single experiment. However, when interested in specific signaling pathways, such shot-gun methodologies lack selectivity, and moreover are not cost and time efficient with respect to instrument and data analysis time. Here we evaluate and explore a peptide-centric antibody generated to selectively enrich peptides containing the cAMP-dependent protein kinase A (PKA) consensus motif. This targeted phospho-proteomic strategy is used to profile temporal quantitative changes of potential protein PKA substrates in Jurkat T-lymphocytes upon prostaglandin E2 stimulation, which increases intracellular cAMP activating PKA. Our method combines ultra-high specificity motif-based immunoaffinity purification with cost-efficient stable isotope dimethyl labeling. We identified 655 phosphopeptides of which 642 (i.e. 98%) contained the consensus motif [R/K][R/K/X]X[pS/pT]. When compared with a large-scale Jurkat T-lymphocytes phosphoproteomics data-set, containing more than 10,500 phosphosites, a minimal overlap of 0.2% was observed. This stresses the need for such targeted analyses when interested in a particular kinase. Our data provides a resource of likely substrates of PKA and potentially some substrates of closely related kinases. Network analysis revealed that about half of the observed substrates have been implicated in cAMP induced signaling. Still, the other half of the here identified substrates have been less well characterized, representing a valuable resource for future research.