A "clickable" MTX reagent as a practical tool for profiling small-molecule-intracellular target interactions via MASPIT.

ABSTRACT: We present a scalable synthesis of a versatile MTX reagent with an azide ligation handle that allows rapid γ-selective conjugation to yield MTX fusion compounds (MFCs) appropriate for MASPIT, a three-hybrid system that enables the identification of mammalian cytosolic proteins that interact with a small molecule of interest. We selected three structurally diverse pharmacologically active compounds (tamoxifen, reversine, and FK506) as model baits. After acetylene functionalization of these baits, MFCs were synthesized via a CuAAC reaction, demonstrating the general applicability of the MTX reagent. In analytical mode, MASPIT was able to give concentration-dependent reporter signals for the established target proteins. Furthermore, we demonstrate that the sensitivity obtained with the new MT

SUBMITTER: Risseeuw MD 

PROVIDER: S-EPMC3790973 | biostudies-literature | 2013 Mar

REPOSITORIES: biostudies-literature