Homogenous fluorescent assays for characterizing small-molecule activators of AMP-activated protein kinase (AMPK).
AMP activated protein kinase (AMPK) is a key regulator of cellular metabolism. AMPK activity is modulated in part by binding of AMP to the gamma-subunit of the kinase, which increases the activity of the catalytic alpha-subunit. Because increased AMPK activity in the liver and in skeletal muscle leads to increased fatty acid oxidation and decreased cholesterol and fatty acid biosynthesis, activators of AMPK are being sought for treatment of type-2 diabetes and other metabolic disorders. The unique mechanism of AMPK activation offers an opportunity to develop small molecules that directly upregulate AMPK activity, and there exists a need for simplified methods to identify and characterize small-molecules that show isoform-specific effects on AMPK. We have developed a suite of fluorescence-based assays to identify and characterize such compounds, and have used these to characterize and compare activity of recombinant AMPK alpha(1)beta(1)gamma(1) and alpha(2)beta(1)gamma(1) isoforms in response to small molecule activators and inhibitors.
Reichling
Laurie J
LJ
Invitrogen Discovery Sciences, 501 Charmany Dr, Madison, WI 53719, USA.
Riddle
Steven M
SM
Mei
Baigen
B
Bruinsma
Rica
R
Goossens
Tony A
TA
Huwiler
Kristin G
KG
Maffitt
Mark
M
Newport
Alyssa M G
AM
Qian
Xiao-Dong
XD
Ruttimann-Johnson
Carmen
C
Vogel
Kurt W
KW