Axon guidance involves the spatiotemporal interplay between guidance cues and membrane-bound cell-surface receptors, present on the growth cone of the axon. Netrin-1 is a prototypical guidance cue that binds to deleted in colorectal cancer (DCC), and it has been proposed that the guidance cue Draxin modulates this interaction. Here, we present structural snapshots of Draxin/DCC and Draxin/Netrin-1 complexes, revealing a triangular relationship that affects Netrin-mediated haptotaxis and fasciculation. Draxin interacts with DCC through the N-terminal four immunoglobulin domains, and Netrin-1 through the EGF-3 domain, in the same region where DCC binds. Netrin-1 and DCC bind to adjacent sites on Draxin, which appears to capture Netrin-1 and tether it to the DCC receptor. We propose the conformational flexibility of the single-pass membrane receptor DCC is used to promote fasciculation and regulate axon guidance through concerted Netrin-1/Draxin binding. Display omitted •Crystal structure of cysteine knot domain of Draxin in complex with DCC•Crystal structure of Netrin-1 in complex with a Draxin fragment•Netrin-1 contains a competing binding site for DCC and Draxin on the EGF-3 domain•Draxin tethers Netrin-1 and DCC together to promote fasciculation Liu et al. report through structural investigations how Draxin associates both with Netrin-1 and its cognate receptor DCC to mediate axon guidance and fasciculation.