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 proteome-wide labeling of bacterial proteins upon activation of the P(BAD) promoter and the SoxRS regulon and provides a basis for analysis of more complex systems including spatially heterogeneous microbial cultures and biofilms.
Division of Chemistry and Chemical Engineering, California Institute of Technology, 1200 E. California Blvd., Pasadena, CA 91125, USA.
John T
JT
Ngo
Brett M
BM
Babin
Julie A
JA
Champion
Erin M
EM
Schuman
David A
DA
Tirrell
D001089:Arabinose / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry; D018441:Biofilms; D002470:Cell Survival; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism; D005456:Fluorescent Dyes / Q000737:chemistry; D005786:Gene Expression Regulation; D005821:Genetic Techniques; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008954:Models, Biological; D008957:Models, Genetic; D011401:Promoter Regions, Genetic; D040901:Proteomics / Q000379:methods; D018085:Regulon; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D008024:Ligands; D001089:Arabinose