In Vivo Reporter Labeling of Proteins via Metabolic Delivery of Coenzyme A Analogues

摘要

Selective chemical control of biochemical processes within a living cell enables the study and modification of natural biological systems in ways that may not be obtained through in vitro experiments.Accordingly,access to promiscuous metabolic pathways has provided a unique chemical entry into small molecule engineering in vivo.We recently introduced a method for covalent reporter labeling of carrier proteins using promiscuous phospho-pantetheinyltransferase(PPTase)enzymes and reporter-labeled coenzyme A(CoA).To date,this method has been limited to in vitro and cell-surface protein labeling,as CoA derivatives have not been shown to penetrate the cell.To overcome this obstacle,we demonstrate addition of labeled metabolic precursors to cell culture that results in cellular uptake and metabolic conversion into active,labeled CoA derivatives.