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

Steven N. Anderson

Department of Biological Screening, Abbott Laboratories, Global Pharmaceutical Research and Development, Abbott Park, IL Steve.n.Anderson{at}abbott.com

Barbara L. Cool

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

Lemma Kifle

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

William Chiou

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

David A. Egan

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

Leo W. Barrett

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

Paul L. Richardson

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

Ernst U. Frevert

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

Usha Warrior

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

James L. Kofron

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

David J. Burns

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

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 (SAMTM). For detection, the SAMTM 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)

Key Words: microarrayed compound screening (µARCS) • high-throughput screening • AMP-activated protein kinase (AMPK) • streptavidin affinity membrane (SAMTM)

Journal of Biomolecular Screening, Vol. 9, No. 2, 112-121 (2004)
DOI: 10.1177/1087057103260592
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