G protein subunit alpha O1 (GNAO1)-related disorders (GNAO1-RD) are a group of ultra-rare neurological conditions characterized by a wide spectrum of clinical features, including movement disorders, developmental delay or intellectual disability, epilepsy, and feeding difficulties. The severity of these conditions can range from mild to severe,1,2 with life-threatening episodes of dyskinetic crisis being a hallmark of the disorder in many cases.3 The Gαo protein, encoded by the GNAO1 gene, is a crucial component of G protein-coupled receptor (GPCR) signaling. It mediates interactions between receptors and intracellular effectors, playing an essential role in neuronal communication and regulation. Structurally, Gαo consists of a Ras-like domain (RHD) and an α-helical domain (AHD), which coordinate nucleotide binding and signaling, along with an N-terminal α-helix (αN). While mutations in the RHD are well-studied due to their role in enzymatic activity and interactions, the functional impact of mutations in the AHD, such as the novel N76K variant of GNAO1, is less understood. Here, we report and analyze the N76K variant, focusing on its effects on Gαo activity and signaling pathways.