Sphingomyelinase‐Mediated Multitimescale Clustering of Ganglioside GM1 in Heterogeneous Lipid Membranes

Several signaling processes in the plasma membrane are intensified by ceramides that are formed by sphingomyelinase‐mediated hydrolysis of sphingomyelin. These ceramides trigger clustering of signaling‐related biomolecules, but how they concentrate such biomolecules remains unclear. Here, the spatiotemporal localization of ganglioside GM1, a glycolipid receptor involved in signaling, during sphingomyelinase‐mediated hydrolysis is described. Real‐time visualization of the dynamic remodeling of the heterogeneous lipid membrane that occurs due to sphingomyelinase action is used to examine GM1 clustering, and unexpectedly, it is found that it is more complex than previously thought. Specifically, lipid membranes generate two distinct types of condensed GM1: 1) rapidly formed but short‐lived GM1 clusters that are formed in ceramide‐rich domains nucleated from the liquid‐disordered phase; and 2) late‐onset yet long‐lasting, high‐density GM1 clusters that are formed in the liquid‐ordered phase. These findings suggest that multiple pathways exist in a plasma membrane to synergistically facilitate the rapid amplification and persistence of signals.