Using the yeast three-hybrid system for the identification of small molecule-protein interactions with the example of ethinylestradiol.

Keywords: ethinylestradiol; lysyl hydroxylase 2; target identification; yeast three-hybrid screen EN ethinylestradiol lysyl hydroxylase 2 target identification yeast three-hybrid screen 421 431 11 03/11/22 20220301 NES 220301 Introduction Many fundamental processes in biology rely on small ligand-protein interactions which form the basis for pharmacological intervention of human diseases. After washing, bound proteins were eluted with SDS and heat, and analyzed by SDS-PAGE and Western Blotting using a PLOD2 antibody for staining, or a GAL4 antibody for the detection of the GAL4 AD fusion protein from the Y3H vector. Having confirmed that our Y3H setup can indeed detect small molecule-protein interactions, we next wanted to employ this system for the identification of protein binding partners of small molecules out of a pool of proteins. [Extracted from the article]