Microarrayed compound screening (microARCS) to identify activators and inhibitors of AMP-activated protein kinase.
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 (microARCS) 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 trade mark ). For detection, the SAM trade mark 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.
Anderson
Steven N
SN
Department of Biological Screening, Abbott Laboratories, Global Pharmaceutical Research and Development, Abbott Park, IL 60064, USA. Steve.n.Anderson@abbott.com
Cool
Barbara L
BL
Kifle
Lemma
L
Chiou
William
W
Egan
David A
DA
Barrett
Leo W
LW
Richardson
Paul L
PL
Frevert
Ernst U
EU
Warrior
Usha
U
Kofron
James L
JL
Burns
David J
DJ
D055372:AMP-Activated Protein Kinases; D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D008904:Miniaturization; D009097:Multienzyme Complexes / Q000032:analysis / Q000037:antagonists & inhibitors*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D017346:Protein Serine-Threonine Kinases / Q000032:analysis / Q000037:antagonists & inhibitors*; D051381:Rats; D012680:Sensitivity and Specificity; D013997:Time Factors
D009097:Multienzyme Complexes; D000255:Adenosine Triphosphate; D017346:Protein Serine-Threonine Kinases; D055372:AMP-Activated Protein Kinases