Journal
JOURNAL OF PROTEOME RESEARCH
Volume 15, Issue 7, Pages 2236-2245Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jproteome.6b00214
Keywords
PePTID; pulse proteolysis; target discovery; energetics-based target discovery approach
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Funding
- National Institute of Science and Technology on Tuberculosis (INCT-TB)
- Decit/SCTIE/MS-MCT-CNPq-FNDCT-CAPES (Brazil)
- FAPERGS
- CNPq [304051/1975-06]
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In recent years, phenotypic screening has assumed a leading role in drug discovery efforts. However, development of new drugs from bioactive compounds obtained in screening campaigns requires identification of the cellular targets responsible for their biological activities. A new energetics-based method for target identification is presented: pulse proteolysis and precipitation for target identification (PePTID). In this method, proteins incubated with or without a ligand and submitted to a brief proteolytic pulse are directly analyzed and compared using a label-free semi quantitative mass spectrometry strategy, dispensing the SDS-PAGE readout and greatly improving the throughput. As a proof-of-concept, we applied the PePTID method to identify ATP binding proteins in Mycobacterium smegmatis, a model system for Mycobacterium tuberculosis, the etiological agent of tuberculosis.
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