State-Selective Metabolic Labeling of Cellular Proteins
Abstract
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.
Additional Information
© 2012 American Chemical Society. Received: May 14, 2012. Accepted: June 7, 2012. Published: June 12, 2012. This work was supported by National Institutes of Health grant NIH R01 GM062523 and by the Institute for Collaborative Biotechnologies through grant W911NF-09-0001 from the U.S. Army Research Office.Attached Files
Published - cb300238w.pdf
Accepted Version - nihms-385141.pdf
Supplemental Material - cb300238w_si_001.pdf
Files
Additional details
- PMCID
- PMC3423470
- Eprint ID
- 35490
- Resolver ID
- CaltechAUTHORS:20121115-132451309
- R01 GM062523
- NIH
- W911NF-09-0001
- Army Research Office (ARO)
- Created
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2012-11-20Created from EPrint's datestamp field
- Updated
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2021-11-09Created from EPrint's last_modified field