Drug Affinity Responsive Target Stability (DARTS) Analysis Service

Drug Affinity Responsive Target Stability (DARTS) technology is an effective method for detecting novel small molecule protein targets.DARTS can be used to validate known small molecule-protein interactions and to find potential protein targets for natural products. When proteins are bound to small molecule compounds, their stability to protein hydrolases is significantly increased. Proteins that bind small molecules are not readily hydrolyzed by proteases. These proteins can then be detected after electrophoresis and subsequently using biomass spectroscopy specific target proteins can be identified.

Drug targets are biological macromolecules that have pharmacodynamic functions within cells and can be acted upon by drugs. Most drug target molecules belong to proteins and are capable of binding small molecule compound drugs with appropriate chemical properties and affinity. After identifying the target molecules relevant to the disease, the relevant drugs can be designed for targeted therapy based on the characteristics of the target to develop effective and safe therapeutic approaches.

For most drugs of small molecules, the key issue is to identify the molecular target, taking into account the therapeutic effect and/or adverse side effects of the drug.

Studies to discover, and identify, biological target proteins within cells through phenotypic screening are becoming increasingly popular. To a large extent, these techniques require certain chemical modifications of small molecule drugs in order to enable them to find target molecules to which they can bind. In addition, some natural product small molecules have limited modification sites due to structural factors, which also constrain the recognition of drug targets by natural products and hinder the research of small molecule drugs and their applications.

Creative Proteomics provides DARTS technology services to overcome these limitations, enabling the analysis of drug-target protein binding relationships and finding the optimal conditions for binding of target proteins to small molecule drugs.

Advantages of DARTS Technology

Universally applicable, simple and fast
Uses the natural, unmodified activity of small molecule drugs with affinity for interaction with target proteins for interaction analysis, rather than using downstream parameters to discover target proteins

DARTS Technology Process

DARTS is a more sensitive technique for protein identification and requires high experimental conditions. The selection and use of key reagent choices such as protease, cell lysate and buffer are all critical factors in determining the success of DARTS. We will specify targeted solutions based on specific samples. A brief process includes:

Cell collection and lysis
Preparation of cell lysate
Incubation of drug with lysate in combination
Proteolytic digestion
Identification of bound proteins: SDS-PAGE, Western protein blotting and LC-MS/MS
By comparing and identifying the significant differences between drug and control lysates, we can find the candidate targets for subsequent experimental confirmation.

Workflow of DARTS (Lomenick et al., 2009)

DARTS technology can identify target proteins and also detect other proteins that have some association with the target protein in the development of disease.

learn more:

molecular docking in drug discovery

drug affinity responsive target stability

Reference

Lomenick, et al. (2009). Target identification using drug affinity responsive target stability (DARTS). Proceedings of the National Academy of Sciences, 106(51), 21984-21989.