Active and inactive orientations of the transmembrane and cytosolic domains of the erythropoietin receptor dimer

Seubert, Nadine [UCL] Royer, Yohan [UCL] Staerk, Judith [UCL] Kubatzky, Katharina F [UCL] Moucadel, Virginie [UCL] Krishnakumar, Shyam Smith, Steven O Constantinescu, Stefan N. [UCL] et al.[show all]

Binding of erythropoietin to the erythropoietin receptor (EpoR) extracellular domain orients the transmembrane (TM) and cytosolic regions of the receptor dimer into an unknown activated conformation. By replacing the EpoR extracellular domain with a dimeric coiled coil, we engineered TM EpoR fusion proteins where the helical TM domains were constrained into seven possible relative orientations. We identify one dimeric TM conformation that imparts full activity to the cytosolic domain of the receptor and signals via JAK2, STAT proteins, and MAP kinase, one partially active orientation that preferentially activates MAP kinase, and one conformation corresponding to the inactive receptor. The active and inactive conformations were independently identified by computational searches for low-energy TM dimeric structures. We propose a specific EpoR-activated interface and suggest its use for structural and signaling studies.