The activity of endothelial nitric oxide synthase (eNOS) is regulated through intricate crosstalk among transcriptional, post-transcriptional, and post-translational modalities. Translational modification cascades and their downstream signaling relay emerge as pivotal controllers of both eNOS functionality and nitric oxide bioavailability, positioning these pathways as prime therapeutic targets. A deeper elucidation of the molecular machinery governing post-translational regulation of eNOS holds promise for developing innovative approaches to forestall endothelial dysfunction and its consequent hypertensive complications.