Structural basis for cell surface patterning through NetrinG-NGL interactions

Brain wiring depends on cells making highly localized and selective connections through surface protein–protein interactions, including those between NetrinGs and NetrinG ligands (NGLs). The NetrinGs are members of the structurally uncharacterized netrin family. We present a comprehensive crystallographic analysis comprising NetrinG1–NGL1 and NetrinG2–NGL2 complexes, unliganded NetrinG2 and NGL3. Cognate NetrinG–NGL interactions depend on three specificity‐conferring NetrinG loops, clasped tightly by matching NGL surfaces. We engineered these NGL surfaces to implant custom‐made affinities for NetrinG1 and NetrinG2. In a cellular patterning assay, we demonstrate that NetrinG‐binding selectivity can direct the sorting of a mixed population of NGLs into discrete cell surface subdomains. These results provide a molecular model for selectivity‐based patterning in a neuronal recognition system, dysregulation of which is associated with severe neuropsychological disorders. NetrinGs and NetrinG‐ligands (NGLs) control neuronal circuit development, and their dysregulation is associated with autism and schizophrenia. The first structures of NetrinG‐NGL complexes and structure‐based functional assays show that binding affinity governs the cell surface compartmentalization of NGLs.