A mechanistic framework for the recognition of chemically diverse brassinosteroids by BRI1-family receptor kinases

Abstract

Brassinosteroids (BRs) are chemically diverse plant steroid hormones produced via a branched biosynthetic pathway. The potent BR brassinolide is sensed by the membrane receptor kinase BRIl and a SERK co-receptor, but the physiological functions of other abundant BRs remain to be characterized. Here, we present quantitative binding kinetics for four Arabidopsis thaliana BR receptors and fifteen BRs, which define the key chemical features required for high affinity receptor binding, ligand positioning, and co-receptor recognition. BRIl, BRLl, and BRL3 share overlapping ligand preferences, whereas BRL2 preferentially binds C28 BRs with moderate affinity. Structural analyses of BR-bound BRIl and BRL3 ectodomains, combined with extensive in vitro and in vivo mutagenesis studies reveal a high structural plasticity of the hormone binding pocket. Functional assays using structure-based BR agonists and antagonists uncover that BR receptor - co-receptor signaling complexes can recognize chemically diverse BRs, introducing an additional, intriguing layer of BR signaling regulation.