Insulin and insulin-like growth factor have an essential role in growth, development and the maintenance of metabolic homeostasis, including glucose uptake from the bloodstream. Researchers have identified mutations in insulin receptors that cause severe insulin resistance1, and a temperature-sensitive daf-2 (a gene encoding an insulin receptor–like protein) mutant in Caenorhabditis elegans has served as an insulin resistance model2. Here we report a forward chemical genetic approach with a tagged library that we used to identify a small molecule, GAPDH segregator (GAPDS), that suppresses the dauer formation induced by the daf-2 mutant. Like insulin, GAPDS increased both glucose uptake and the concentration of phosphatidylinositol-3,4,5-trisphosphate (PtdIns(3,4,5)P3) in mammalian preadipocytes. Using affinity matrices and RNA interference, we identified glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a GAPDS target. We discovered that GAPDH stimulates phosphatase activity against not only PtdIns(3,4,5)P3 but also PtdIns(4,5)P2. These results suggest that GAPDH is both an active regulator in the phosphoinositide-mediated signaling pathway and a potential new target for insulin resistance treatment.
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We gratefully acknowledge the support of the US National Institutes of Health (NIH) (R01-CA096912), and the US National Science Foundation equipment grants for the NMR (MRI-0116222) and the capillary LC ion trap mass spectrometer (CHE-0234863). Components of this work were conducted in a Shared Instrumentation Facility constructed with support from Research Facilities Improvement grant C06 RR-16572 from the US National Center for Research Resources and the US NIH. J.R.Y. is supported by US NIH grant P41RR11823-10.
The authors declare no competing financial interests.
Synthetic scheme and structure of TG-Bz library. (PDF 42 kb)
DAF-2 signaling pathway and research strategy for the forward chemical genetic study using daf-2 mutant in C. elegans model. (PDF 135 kb)
The effect of compounds on protein cross-linking. (PDF 30 kb)
GAPDS effect on GAPDH in 3T3-L1 cell lysate. (PDF 114 kb)
GAPDH siRNA effect on cellular PIP2 level. (PDF 35 kb)
Target identification using chemical affinity matrix and RNAi. (PDF 15 kb)
Experimental procedures for the synthesis of the TG-Bz library, immobilization method for preparing GAPDS affinity matrix, in vitro GAPDH inhibition assay, and statistical analysis of the data. (PDF 80 kb)
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Min, J., Kyung Kim, Y., Cipriani, P. et al. Forward chemical genetic approach identifies new role for GAPDH in insulin signaling. Nat Chem Biol 3, 55–59 (2007). https://doi.org/10.1038/nchembio833
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