State-selective Metabolic Labeling of Cellular Proteins

Transcriptional activity from a specified promoter can provide a useful marker for the physiological state of a cell. Here we introduce a method for selective tagging of proteins made in cells in which specified promoters are active. Tagged proteins can be modified with affinity reagents for enrichment or with fluorescent dyes for visualization. The method allows state-selective analysis of the proteome, whereby proteins synthesized in predetermined physiological states can be identified. The approach is demonstrated by proteorne-wide labeling of bacterial proteins upon activation of the P-BAD promoter and the SoocRS regulon and provides a basis for analysis of more complex systems including spatially heterogeneous microbial cultures and biofilms.