Chemical Synthesis of Phosphorylated Ubiquitin and Diubiquitin Exposes Positional Sensitivities of E1‐E2 Enzymes and Deubiquitinases |
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Authors: | Dr. Somasekhar Bondalapati Wissam Mansour Dr. Mark A. Nakasone Dr. Suman Kumar Maity Prof. Michael H. Glickman Prof. Ashraf Brik |
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Affiliation: | 1. Department of Chemistry, Ben‐Gurion University of the Negev, P.O. Box 653 Beer‐Sheva 8410501 (Israel);2. Schulich Faculty of Chemistry, Technion‐Israel Institute of Technology, 3200008 Haifa (Israel);3. Department of Biology, Technion‐Israel Institute of Technology, 3200008 Haifa (Israel) |
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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. |
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Keywords: | deubiquitinases ligases phosphorylation ubiquitin chains ubiquitination |
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