Microarrayed Compound Screening (μARCS) to Identify Activators and Inhibitors of AMP-Activated Protein Kinase

Author:

Anderson Steven N.1,Cool Barbara L.2,Kifle Lemma2,Chiou William2,Egan David A.3,Barrett Leo W.4,Richardson Paul L.4,Frevert Ernst U.2,Warrior Usha1,Kofron James L.1,Burns David J.1

Affiliation:

1. Department of Biological Screening, Abbott Laboratories, Global Pharmaceutical Research and Development, Abbott Park, IL

2. Department of Metabolic Disease Research, Abbott Laboratories, Global Pharmaceutical Research and Development, Abbott Park, IL

3. Department of Protein Biochemistry, Abbott Laboratories, Global Pharmaceutical Research and Development, Abbott Park, IL

4. Department of Structural Biology, Abbott Laboratories, Global Pharmaceutical Research and Development, Abbott Park, IL

Abstract

A novel and innovative high-throughput screening assay was developed to identify both activators and inhibitors of AMP-activated protein kinase (AMPK) using microarrayed compound screening (μARCS) technology. Test compounds were arrayed at a density of 8640 on a polystyrene sheet, and the enzyme and peptide substrate were introduced into the assay by incorporating them into an agarose gel followed by placement of the gels onto the compound sheet. Adenosine triphosphate (ATP) was delivered via a membrane, and the phosphorylated biotinylated substrate was captured onto a streptavidin affinity membrane (SAM™). For detection, the SAM™ was removed, washed, and imaged on a phosphor screen overnight. A library of more than 700,000 compounds was screened using this format to identify novel activators and inhibitors of AMPK. ( Journal of Biomolecular Screening 2004:112-121)

Publisher

Elsevier BV

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