title: Synthesis and Use of a Phosphonate Amidine to Generate an Anti-Phosphoarginine-Specific Antibody.
abstract: Protein arginine phosphorylation is a post-translational modification (PTM) that is important for bacterial growth and virulence. Despite its biological relevance, the intrinsic acid lability of phosphoarginine (pArg) has impaired studies of this novel PTM. Herein, we report for the first time the development of phosphonate amidines and sulfonate amidines as isosteres of pArg and then use these mimics as haptens to develop the first high-affinity sequence independent anti-pArg specific antibody. Employing this anti-pArg antibody, we further showed that arginine phosphorylation is induced in Bacillus subtilis during oxidative stress. Overall, we expect this antibody to see widespread use in analyzing the biological significance of arginine phosphorylation. Additionally, the chemistry reported here will facilitate the generation of pArg mimetics as highly potent inhibitors of the enzymes that catalyze arginine phosphorylation/dephosphorylation.
authors.affiliation: Department of Chemistry, The Scripps Research Institute, 130 Scripps Way, Jupiter, FL 33458 (USA). jfuhrman@scripps.edu.
authors.forename: Jakob
authors.initials: J
authors.lastname: Fuhrmann
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, UMass Medical School, 364 Plantation Street, Worcester, MA 01605 (USA).|Chemical Biology Interface Program, UMass Medical School, 364 Plantation Street, Worcester, MA 01605 (USA).
authors.forename: Venkataraman
authors.initials: V
authors.lastname: Subramanian
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, UMass Medical School, 364 Plantation Street, Worcester, MA 01605 (USA). paul.thompson@umassmed.edu.|Chemical Biology Interface Program, UMass Medical School, 364 Plantation Street, Worcester, MA 01605 (USA). paul.thompson@umassmed.edu.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Thompson
mesh_terms: D000578:Amidines / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D001120:Arginine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D001412:Bacillus subtilis / Q000378:metabolism; D006241:Haptens / Q000737:chemistry / Q000276:immunology; D015394:Molecular Structure; D063065:Organophosphonates / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D009943:Organophosphorus Compounds / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D018384:Oxidative Stress; D010766:Phosphorylation
chemical_list: D000578:Amidines; D000906:Antibodies; D006241:Haptens; D063065:Organophosphonates; D009943:Organophosphorus Compounds; C015441:phospho-L-arginine; D001120:Arginine
keywords: N-phosphorylation; antibodies; haptens; phosphoarginine; phosphonate amidine