Differential radial capillary action of ligand assay for high-throughput detection of protein-metabolite interactions.

ABSTRACT: Interactions of proteins with low-molecular-weight ligands, such as metabolites, cofactors, and allosteric regulators, are important determinants of metabolism, gene regulation, and cellular homeostasis. Pharmaceuticals often target these interactions to interfere with regulatory pathways. We have developed a rapid, precise, and high-throughput method for quantitatively measuring protein-ligand interactions without the need to purify the protein when performed in cells with low background activity. This method, differential radial capillary action of ligand assay (DRaCALA), is based on the ability of dry nitrocellulose to separate the free ligand from bound protein-ligand complexes. Nitrocellulose sequesters proteins and bound ligand at the site of application, whereas free ligand is mobil

SUBMITTER: Roelofs KG 

PROVIDER: S-EPMC3174574 | biostudies-literature | 2011 Sep

REPOSITORIES: biostudies-literature