title: Chemical synthesis of phosphorylated ubiquitin and diubiquitin exposes positional sensitivities of e1-e2 enzymes and deubiquitinases.
abstract: Modification of ubiquitin by phosphorylation extends the signaling possibilities of this dynamic signal, as it could affect the activity of ligases and the processing of ubiquitin chains by deubiquitinases. The first chemical synthesis of phosphorylated ubiquitin and of Lys63-linked diubiquitin at the proximal, distal or both ubiquitins is reported. This enabled the examination of how such a modification alters E1-E2 activities of the ubiquitination machinery. It is found that E1 charging was not affected, while the assembly of phosphorylated ubiquitin chains was differentially inhibited with E2 enzymes tested. Moreover, this study shows that phosphorylation interferes with the recognition of linkage specific antibodies and the activities of several deubiquitinases. Notably, phosphorylation in the proximal or distal ubiquitin unit has differential effects on specific deubiquitinases. These results support a unique role of phosphorylation in the dynamics of the ubiquitin signal.
authors.affiliation: Department of Chemistry, Ben-Gurion University of the Negev, P.O. Box 653 Beer-Sheva 8410501 (Israel); Schulich Faculty of Chemistry, Technion-Israel Institute of Technology, 3200008 Haifa (Israel).
authors.forename: Somasekhar
authors.initials: S
authors.lastname: Bondalapati
authors.forename: Wissam
authors.initials: W
authors.lastname: Mansour
authors.forename: Mark A
authors.initials: MA
authors.lastname: Nakasone
authors.forename: Suman Kumar
authors.initials: SK
authors.lastname: Maity
authors.forename: Michael H
authors.initials: MH
authors.lastname: Glickman
authors.forename: Ashraf
authors.initials: A
authors.lastname: Brik
mesh_terms: D008025:Ligases / Q000737:chemistry* / Q000378:metabolism; D010766:Phosphorylation; D025801:Ubiquitin / Q000138:chemical synthesis* / Q000737:chemistry*; D064570:Ubiquitin-Specific Proteases / Q000737:chemistry* / Q000378:metabolism
chemical_list: D025801:Ubiquitin; D064570:Ubiquitin-Specific Proteases; D008025:Ligases
keywords: deubiquitinases; ligases; phosphorylation; ubiquitin chains; ubiquitination