A targeted approach for the synthesis of multi-phosphorylated peptides: a tool for studying the role of phosphorylation patterns in proteins. Issue 42 (9th September 2019)

• Record Type: Journal Article
• Authors: Samarasimhareddy, Mamidi; Mayer, Daniel; Metanis, Norman; Veprintsev, Dmitry; Hurevich, Mattan; Friedler, Assaf
• Is Part Of: Organic & biomolecular chemistry. Volume 17:Issue 42(2019)
• Page Start: 9284
• Page End: 9290
• Publication Date: 2019-09-09
• Subjects: Chemistry, Organic -- Periodicals; Bioorganic chemistry -- Periodicals; Chemistry, Physical organic -- Periodicals; 547
• DOI: 10.1039/c9ob01874c
• Languages: English
• ISSNs: 1477-0520

Abstract

Abstract : An efficient strategy for the synthesis of multiphosphorylated peptides is described. Such peptides are essential in elucidating the biological roles of the phosphorylation patterns and barcodes. Abstract : Protein phosphorylation barcodes, clusters of several phosphorylation sites within a short unfolded region, control many cellular processes. Existing biochemical methods used to study the roles of these barcodes suffer from low selectivity and provide only qualitative data. Chemically synthesized multiphosphopeptide libraries are selective and specific, but their synthesis is extremely difficult using the current peptide synthesis methods. Here we describe a new microwave assisted approach for synthesizing a library of multiphosphopeptides, using the C-terminus of rhodopsin as a proof of concept. Our approach utilizes multiple protocols for synthesizing libraries of multiphosphopeptides instead of the inefficient single protocol methods currently used. Using our approach we demonstrated the synthesis with up to seven phosphorylated amino acids, sometimes next to each other, an accomplishment that was impractical before. Synthesizing the Rhodopsin derived multiphosphopeptide library enabled dissecting the precise phosphorylation barcode required for the recruitment, activation and modulation of the conformation of Arrestin. Since phosphorylation barcodes modulate the activity of hundreds of GPCRs, synthesizing libraries of multiphosphopeptides is the method of choice for studying their molecular mechanisms of action. Our approach provides an invaluable tool for evaluating how protein phosphorylation barcodes regulate their activity.