title: Structural basis for recognizing phosphoarginine and evolving residue-specific protein phosphatases in gram-positive bacteria.
abstract: Many cellular pathways are regulated by the competing activity of protein kinases and phosphatases. The recent identification of arginine phosphorylation as a protein modification in bacteria prompted us to analyze the molecular basis of targeting phospho-arginine. In this work, we characterize an annotated tyrosine phosphatase, YwlE, that counteracts the protein arginine kinase McsB. Strikingly, structural studies of YwlE reaction intermediates provide a direct view on a captured arginine residue. Together with biochemical data, the crystal structures depict the evolution of a highly specific phospho-arginine phosphatase, with the use of a size-and-polarity filter for distinguishing phosphorylated arginine from other phosphorylated side chains. To confirm the proposed mechanism, we performed bioinformatic searches for phosphatases, employing a similar selectivity filter, and identified a protein in Drosophila melanogaster exhibiting robust arginine phosphatase activity. In sum, our findings uncover the molecular framework for specific targeting of phospho-arginine and suggest that protein arginine (de)phosphorylation may be relevant in eukaryotes.
authors.affiliation: Research Institute of Molecular Pathology (IMP), A-1030 Vienna, Austria.
authors.forename: Jakob
authors.initials: J
authors.lastname: Fuhrmann
authors.forename: Beata
authors.initials: B
authors.lastname: Mierzwa
authors.forename: Débora B
authors.initials: DB
authors.lastname: Trentini
authors.forename: Silvia
authors.initials: S
authors.lastname: Spiess
authors.forename: Anita
authors.initials: A
authors.lastname: Lehner
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Charpentier
authors.forename: Tim
authors.initials: T
authors.lastname: Clausen
mesh_terms: D000595:Amino Acid Sequence; D001120:Arginine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006094:Gram-Positive Bacteria / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D009943:Organophosphorus Compounds / Q000737:chemistry / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D009943:Organophosphorus Compounds; C015441:phospho-L-arginine; D001120:Arginine; D010749:Phosphoprotein Phosphatases