Pulse Proteolysis and Precipitation for Target Identification.

Abstract

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 semiquantitative 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.

• Authors: Rogério V Trindade, Antônio F M Pinto, Diógenes S Santos, Cristiano V Bizarro
• Journal: Journal of proteome research (J Proteome Res) Vol. 15 Issue 7 Pg. 2236-45 (07 01 2016) ISSN: 1535-3907 [Electronic] United States
• PMID: 27255303 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• ATP-binding protein, bacteria
• Bacterial Proteins
• Carrier Proteins
• Ligands

Topics

• Bacterial Proteins (analysis)
• Carrier Proteins (analysis)
• Chemical Precipitation
• Drug Discovery (methods)
• Ligands
• Mycobacterium smegmatis (chemistry)
• Mycobacterium tuberculosis (chemistry)
• Proteolysis