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An autophosphorylation site database for leucine‐rich repeat receptor‐like kinases in A rabidopsis thaliana
Author(s) -
Mitra Srijeet K.,
Chen Ruiqiang,
Dhandaydham Murali,
Wang Xiaofeng,
Blackburn Robert Kevin,
Kota Uma,
Goshe Michael B.,
Schwartz Daniel,
Huber Steven C.,
Clouse Steven D.
Publication year - 2015
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/tpj.12863
Subject(s) - autophosphorylation , biology , phosphorylation , protein kinase domain , leucine rich repeat , kinase , arabidopsis , arabidopsis thaliana , microbiology and biotechnology , genetics , protein kinase a , mutant , gene
Summary Leucine‐rich repeat receptor‐like kinases ( LRR RLK s) form a large family of plant signaling proteins consisting of an extracellular domain connected by a single‐pass transmembrane sequence to a cytoplasmic kinase domain. Autophosphorylation on specific Ser and/or Thr residues in the cytoplasmic domain is often critical for the activation of several LRR RLK family members with proven functional roles in plant growth regulation, morphogenesis, disease resistance, and stress responses. While identification and functional characterization of in vivo phosphorylation sites is ultimately required for a full understanding of LRR RLK biology and function, bacterial expression of recombinant LRR RLK cytoplasmic catalytic domains for identification of in vitro autophosphorylation sites provides a useful resource for further targeted identification and functional analysis of in vivo sites. In this study we employed high‐throughput cloning and a variety of mass spectrometry approaches to generate an autophosphorylation site database representative of more than 30% of the approximately 223 LRR RLK s in Arabidopsis thaliana . We used His‐tagged constructs of complete cytoplasmic domains to identify a total of 592 phosphorylation events across 73 LRR RLK s, with 497 sites uniquely assigned to specific Ser (268 sites) or Thr (229 sites) residues in 68 LRR RLK s. Multiple autophosphorylation sites per LRR RLK were the norm, with an average of seven sites per cytoplasmic domain, while some proteins showed more than 20 unique autophosphorylation sites. The database was used to analyze trends in the localization of phosphorylation sites across cytoplasmic kinase subdomains and to derive a statistically significant sequence motif for phospho‐Ser autophosphorylation.