title: Phosphoproteomic analysis reveals major default phosphorylation sites outside long intrinsically disordered regions of Arabidopsis plasma membrane proteins.
abstract: BACKGROUND
  Genome-wide statistics established that long intrinsically disordered regions (over 30 residues) are predicted in a large part of proteins in all eukaryotes, with a higher ratio in trans-membrane proteins. At functional level, such unstructured and flexible regions were suggested for years to favour phosphorylation events. In plants, despite increasing evidence of the regulation of transport and signalling processes by phosphorylation events, only few data are available without specific information regarding plasma membrane proteins, especially at proteome scale.
  
  
  RESULTS
  Using a dedicated phosphoproteomic workflow, 75 novel and unambiguous phosphorylation sites were identified in Arabidopsis plasma membrane. Bioinformatics analysis showed that this new dataset concerned mostly integral proteins involved in key functions of the plasma membrane (such as transport and signal transduction, including protein phosphorylation). It thus expanded by 15% the directory of phosphosites previously characterized in signalling and transport proteins. Unexpectedly, 66% of phosphorylation sites were predicted to be located outside long intrinsically disordered regions. This result was further corroborated by analysis of publicly available data for the plasma membrane.
  
  
  CONCLUSIONS
  The new phosphoproteomics data presented here, with published datasets and functional annotation, suggest a previously unexpected topology of phosphorylation in the plant plasma membrane proteins. The significance of these new insights into the so far overlooked properties of the plant plasma membrane phosphoproteome and the long disordered regions is discussed.
authors.affiliation: UR1199 Laboratoire de Protéomique Fonctionnelle, INRA, 34060, Montpellier cedex, France. nespoulo@supagro.inra.fr.
authors.forename: Claude
authors.initials: C
authors.lastname: Nespoulous
authors.forename: Valérie
authors.initials: V
authors.lastname: Rofidal
authors.forename: Nicolas
authors.initials: N
authors.lastname: Sommerer
authors.forename: Sonia
authors.initials: S
authors.lastname: Hem
authors.forename: Michel
authors.initials: M
authors.lastname: Rossignol