Structural basis of death domain signaling in the p75 neurotrophin receptor

Death domains (DDs) mediate assembly of oligomeric complexes for activation of downstream signaling pathways through incompletely understood mechanisms. Here we report structures of complexes formed by the DD of p75 neurotrophin receptor (p75NTR) with RhoGDI, for activation of the RhoA pathway, with caspase recruitment domain (CARD) of RIP2 kinase, for activation of the NF-kB pathway, and with itself, revealing how DD dimerization controls access of intracellular effectors to the receptor. RIP2 CARD and RhoGDI bind to p75NTR DD at partially overlapping epitopes with over 100-fold difference in affinity, revealing the mechanism by which RIP2 recruitment displaces RhoGDI upon ligand binding. The p75NTR DD forms non-covalent, low-affinity symmetric dimers in solution. The dimer interface overlaps with RIP2 CARD but not RhoGDI binding sites, supporting a model of receptor activation triggered by separation of DDs. These structures reveal how competitive protein-protein interactions orchestrate the hierarchical activation of downstream pathways in non-catalytic receptors. DOI: http://dx.doi.org/10.7554/eLife.11692.001
eLife digest Cells have proteins called receptors on their surface that can bind to specific molecules on the outside of the cell. Typically, this binding activates the receptor and the activated receptor then triggers some biochemical changes inside the cell. For many receptors, the portion of the receptor inside the cell is essentially an enzyme that can trigger a biochemical change by itself. Some receptors, however, lack any enzymatic activity, and it is often unclear how these ‘non-catalytic receptors’ trigger changes inside a cell. A protein called p75 neurotrophin receptor (or p75NTR for short) is a non-catalytic receptor that is expressed when neurons are injured and its activity leads to the death of the neurons and related cells. Inhibiting this non-catalytic receptor is an attractive strategy for limiting the damage caused by diseases of the nervous system. However, the molecular mechanisms behind the activity of p75NTR are not well understood. Previous biochemical studies set out to answer the question of how p75NTR engages with components of the signaling machinery inside the cell, and found several components that interact with this receptor. Now, Lin et al. have tried to gain a more detailed understanding of those interactions at a molecular level. This involved solving the three-dimensional structures of three protein complexes that involve part of p75NTR (called the “death domain”) and one of two signaling components (called RhoGDI and RIP2). Two of the protein complexes showed that RIP2 and RhoGDI bind to the receptor’s death domain at partially overlapping sites, although RIP2 binds about 100 times more strongly than RhoGDI.A third protein complex showed an interaction between two copies of the death domain, which involves a surface of the receptor that overlaps with RIP2’s, but not RhoGDI’s, binding site. These structures, together with the results of other experiments, allowed Lin et al. to propose a model that could explain how p75NTR is activated. First, the two death domains must be separated. Next, RIP2 is recruited to the receptor, and outcompetes and displaces RhoGDI. This change in protein-protein interactions switches the receptor’s signaling from one pathway to the other. Now that these structures are available, they can be used in future experiments to design specific changes in the receptor that would allow researchers to dissect its different activities. DOI: http://dx.doi.org/10.7554/eLife.11692.002