title: A proteomic approach identifies many novel palmitoylated proteins in Arabidopsis.
abstract: S-acylation (palmitoylation) is a poorly understood post-translational modification of proteins involving the addition of acyl lipids to cysteine residues. S-acylation promotes the association of proteins with membranes and influences protein stability, microdomain partitioning, membrane targeting and activation state. No consensus motif for S-acylation exists and it therefore requires empirical identification. Here, we describe a biotin switch isobaric tagging for relative and absolute quantification (iTRAQ)-based method to identify S-acylated proteins from Arabidopsis. We use these data to predict and confirm S-acylation of proteins not in our dataset. We identified c. 600 putative S-acylated proteins affecting diverse cellular processes. These included proteins involved in pathogen perception and response, mitogen-activated protein kinases (MAPKs), leucine-rich repeat receptor-like kinases (LRR-RLKs) and RLK superfamily members, integral membrane transporters, ATPases, soluble N-ethylmaleimide-sensitive factor-activating protein receptors (SNAREs) and heterotrimeric G-proteins. The prediction of S-acylation of related proteins was demonstrated by the identification and confirmation of S-acylation sites within the SNARE and LRR-RLK families. We showed that S-acylation of the LRR-RLK FLS2 is required for a full response to elicitation by the flagellin derived peptide flg22, but is not required for localization to the plasma membrane. Arabidopsis contains many more S-acylated proteins than previously thought. These data can be used to identify S-acylation sites in related proteins. We also demonstrated that S-acylation is required for full LRR-RLK function.
authors.affiliation: School of Biological Science, University of Bristol, Woodland Road, Bristol, BS8 1UG, UK.
authors.forename: Piers A
authors.initials: PA
authors.lastname: Hemsley
authors.affiliation: Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QW, UK.
authors.forename: Thilo
authors.initials: T
authors.lastname: Weimar
authors.affiliation: Cambridge Centre for Proteomics, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QR, UK.
authors.forename: Kathryn S
authors.initials: KS
authors.lastname: Lilley
authors.affiliation: Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QW, UK.
authors.forename: Paul
authors.initials: P
authors.lastname: Dupree
authors.affiliation: School of Biological Science, University of Bristol, Woodland Road, Bristol, BS8 1UG, UK.
authors.forename: Claire S
authors.initials: CS
authors.lastname: Grierson
mesh_terms: D000215:Acylation; D000217:Acyltransferases / Q000378:metabolism; D017360:Arabidopsis / Q000737:chemistry / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D001665:Binding Sites; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods*; D050600:SNARE Proteins / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D029681:Arabidopsis Proteins; D050600:SNARE Proteins; D000217:Acyltransferases; C506684:TIP1 protein, Arabidopsis