A Bifunctional Noncanonical Amino Acid: Synthesis, Expression, and Residue-Specific Proteome-wide Incorporation.
Fiche publication
Date publication
août 2018
Journal
Biochemistry
Auteurs
Membres identifiés du Cancéropôle Est :
Dr DZIUBA Dmytro
Tous les auteurs :
Hoffmann JE, Dziuba D, Stein F, Schultz C
Lien Pubmed
Résumé
Mapping of weak and hence transient interactions between low-abundance interacting molecules is still a major challenge in systems biology and protein biochemistry. Therefore, additional system-wide acting tools are needed to determine protein interactomics. Most important are reagents that can be applied at any kind of protein interface and the possibility to enrich cross-linked fragments with high efficiency. In this study, we report the synthesis of a novel noncanonical amino acid that features a diazirine group for ultraviolet cross-linking as well as an alkyne group for labeling by click chemistry. This bifunctional amino acid, called PrDiAzK, may be inserted into almost any protein interface with minimal structural perturbation using genetic code expansion. We demonstrate that PrDiAzK can be site-selectively incorporated into proteins in both bacterial and mammalian cell cultures, and we show that PrDiAzK allows protein labeling as well as cross-linking. In addition, we tested PrDiAzK for proteome-wide incorporation via stochastic orthogonal recoding of translation, implying potential applications in system-wide mapping of protein-protein interactions in the future.
Mots clés
Amino Acids, chemical synthesis, Animals, Click Chemistry, methods, Proteome, analysis
Référence
Biochemistry. 2018 08 7;57(31):4747-4752