1. Proof of concept: Pull down assay using bovine serum albumin and an immunomodulator small molecule
- Author
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Yasser Tabana, Chih-Hsuan Lin, Dinesh Babu, Ramanaguru Siva‐Piragasam, Ashley A. Ponich, Tae Chul Moon, Arno G. Siraki, Shokrollah Elahi, Richard Fahlman, Frederick G. West, and Khaled Barakat
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Science (General) ,Q1-390 ,Social sciences (General) ,H1-99 - Abstract
In the past decade, there has been increasing interest in use of small molecules for immunomodulation. The affinity-based pull-down purification is an essential tool for target identification of small molecules and drug discovery. This study presents our recent efforts to investigate the cellular target(s) of Compound A, a small molecule with demonstrated immunomodulatory properties in human peripheral blood mononuclear cells (PBMCs). While we have previously observed the immunomodulatory activity of Compound A in PBMCs, the specific molecular targets underlying its effects remains elusive. To address this challenge, we synthesized a trifluoromethyl phenyl diazirine (TPD)-bearing trifunctional Probe 1 based on the chemical structure of Compound A, which could be used in a pull-down assay to efficiently bind to putative cellular targets via photoaffinity labelling. In this report, we utilized bovine serum albumin (BSA) as a model protein to establish a proof-of-concept in order to assess the suitability of Probe 1 for binding to an endogenous target. By the successful synthesis of Probe 1 and demonstrating the efficient binding of Probe 1 to BSA, we propose that this method can be used as a tool for further identification of potential protein targets of small molecules in living cells. Our findings provide a valuable starting point for further investigations into the molecular mechanisms underlying the immunomodulatory effects of Compound A.
- Published
- 2023
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