Generation of nucleotide-linked resins for identification of novel binding proteins

Abstract

Organisms use numerous nucleotide-containing compounds as intracellular signals to control behavior. Identifying the biomolecules responsible to sensing and responding to changes in signaling molecule concentration is an important area of research. However, identifying the binding proteins can be challenging when there is no prior information available about binding motifs. In this chapter, we describe a straightforward method to generate nucleotide-linked resins for use in pull-down experiments to identify binding proteins. The protocol outlined in this chapter also can be adapted to generate custom resins linked to other molecules of interest.

Keywords: 2′,3′-cyclic nucleotide monophosphate; Binding proteins; Cyclic nucleotide; Nucleotide coupling; Nucleotide resin; Nucleotide signaling; Protein pull-down; Pull-down.