An Orthogonal and Reversible Protein Labeling via Phosphopantetheinylation

University Of California, San Diego, La Jolla CA

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Abstract

? DESCRIPTION (provided by applicant): The advancement of protein labeling methodology is key to accelerating high-content microscopy technologies, expediting proteomic analyses, and expanding our tools to identify and monitor disease. This program examines the development of a reversible system to label selected proteins. Unlike conventional approaches, the proposed system adapts peptide-screening technologies as a tool to encode discrete selectivity within a two-stage enzymatic pair reaction. Selective labeling is dictated by the choice of labeling enzymes. The proposed program begins through the rational development of orthogonal tags by computational analysis. Evaluation of this system includes detailed kinetic analyses and applications for understanding protein complexes. Overall, this system opens new avenues in protein science. This program will provide a new method for rapidly labeling specific proteins within the complex compendium of proteins towards future studies within the cell or tissue.