Open AccessTarget Identification Using Drug Affinity Responsive Target Stability (DARTS)
Author(s) -
Lomenick Brett,
Jung Gwanghyun,
Wohlschlegel James A.,
Huang Jing
Publication year - 2011
Publication title -
current protocols in chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.503
H-Index - 14
ISSN - 2160-4762
DOI - 10.1002/9780470559277.ch110180
Subject(s) - small molecule , ligand (biochemistry) , target protein , protein ligand , computational biology , protein–protein interaction , chemistry , identification (biology) , protease , drug , combinatorial chemistry , biophysics , biological system , biology , biochemistry , receptor , enzyme , pharmacology , botany , gene
Abstract Drug affinity responsive target stability (DARTS) is a general methodology for identifying and studying protein‐ligand interactions. The technique is based on the principle that when a small molecule compound binds to a protein, the interaction stabilizes the target protein's structure such that it becomes resistant to proteases. DARTS is particularly useful for the initial identification of the protein targets of small molecules, but can also be used to validate potential protein‐ligand interactions predicted or identified by other means and to estimate the affinity of interactions. The approach is simple and advantageous because it can be performed using crude cell lysates and other complex protein mixtures (without requiring purified proteins), and it uses native, unmodified small molecules. The protocols in this unit describe the general approach for performing DARTS experiments, which can be easily modified and scaled to fit project‐specific criteria. Curr. Protoc. Chem. Biol . 3:163‐180 © 2011 by John Wiley & Sons, Inc.