Analyses of the sequence and nature of all protein amino-termini (N-termini) within proteomes (N-terminome) in eukaryotes reveal a highly underestimated occurrence of database nonannotated alternative translates, thereby increasing the already high complexity of proteomes. Nterminomics provides quantitative information on the N-terminal modification states of identified proteins. Ribosome profiling, a recently developed genomics method, enables systematic monitoring of protein translation events by deep sequencing of ribosome-protected mRNA fragments and thereby enables the study of (alternative) translation (initiation) with subcodon resolution. Combining N-terminomics with ribosome profiling will permit the delineation of various dynamic parameters, including translation efficiencies and the occurrence of translation stalling, which might reveal in depth information about the yet incompletely understood quantitative nature of translation initiation and its associated co-translational N-terminal protein modifications (e.g., protein N-terminal acetylation). Bioinformatics-assisted integration of mass spectrometry and ribosome profiling data is here presented to attest its potential for (re- )annotating the translation initiation landscape in human in order to study its dynamicity and regulation. Further, selective ribosome profiling will be used to study the substrate specificity and interplay of ribosome-associated nascent chain interacting factors at the genome level.