Discovery of a sulfotyrosine-motif in the human TrkB extracellular domain required for agonist activation

The brain-derived neurotrophic factor (BDNF)-tropomyosin receptor kinase B (TrkB) signalling axis is a key effector of synaptic plasticity and neuroprotection. While TrkB activation is a major objective towards preventing dysfunction of the nervous system, it cannot be reached with exogenous BDNF administration given the unfavourable physiochemical properties of BDNF. In addition, BDNF also activates a tumour necrosis factor pathway by binding to the neurotrophin receptor p75. The TrkB agonist ZEB85 provides an alternative route to the selective activation of TrkB. We report here the structural basis for the interaction between human TrkB, and both ZEB85 and BDNF, and reveal that a sulfated tyrosine modification is indispensable for ZEB85 activation of TrkB signalling. Using structure-guided BDNF- and ZEB85-binding deficient TrkB mutants, we assessed their ability to sequester ligands from full-length TrkB in cultured human neurons. We found that the BDNF binding site extends into the extracellular juxtamembrane domain of TrkB but does not require the sulfotyrosine at residue 400 to activate TrkB. Together with biophysical analysis and AlphaFold modelling these results also explain how BDNF can displace ZEB85 from TrkB through an overlapping epitope. Our findings reveal unique features of TrkB, not present in the related neurotrophin receptors TrkA and TrkC, and suggest new directions to explore the role of sulfotyrosine in TrkB signalling and identify new TrkB-specific protein ligands. One Sentence SummaryInvestigation of the mechanism of action of TrkB agonist ZEB85 extends molecular understanding of TrkB activation.