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Updated: Apr 11, 2026

2 in 1: One-step Affinity Purification for the Parallel Analysis of Protein-Protein and Protein-Metabolite Complexes
Published on: August 6, 2018
Genetic Diversity, Predictive Protein Structures, and Interaction Networks of Cysteine-Rich Receptor-Like Kinases in
Jente Stouthamer1, Danilo Pereira1, Sergio Martin-Ramirez1
1Laboratory of Biochemistry, Wageningen University and Research, 6708 WE Wageningen, The Netherlands.
Abstract:
Cysteine-rich receptor-like kinases (CRKs) are a large subfamily of plant receptor-like kinases implicated in immunity and development, yet their ligands, interaction partners, and mechanistic roles remain poorly defined. We combined population-genetic analyses and AlphaFold-based structural prediction to characterize the Arabidopsis thaliana CRK family. Phylogenetic reconstruction from 69 natural accessions resolved 5 well-supported CRK clades. Nucleotide diversity (π) and neutrality tests revealed heterogeneous diversity across loci, with evidence of both positive and negative selection pressure acting on different CRKs. AlphaFold models of CRK extracellular domains (ECDs) recapitulate the DUF26 structure observed in plasmodesmata localizing protein (PDLP)5/PDLP8 and ginkbilobin-2 but display distinct biochemical properties and disulfide-bond topologies. Pairwise AlphaFold dimer modeling of all 780 CRK-ECD combinations produced 145 high-confidence interaction models; ~78% of these adopt a shared dimer conformation characterized by an extended intermolecular β-sheet at the interface. Integrating evolutionary and structural approaches reveal clade-specific selective regimes and conserved structural features of CRK-ECDs that likely underpin receptor-receptor interactions. Predicted high-confidence dimer interfaces suggest a general mode of CRK-ECD association that can guide targeted biochemical and genetic validation, accelerating functional dissection of this important receptor family.
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