Insulin and oleate promote translocation of inosine-5' monophosphate dehydrogenase to lipid bodies.
In the present study we identify inosine-5' monophosphate dehydrogenase (IMPDH), a key enzyme in de novo guanine nucleotide biosynthesis, as a novel lipid body-associated protein. To identify new targets of insulin we performed a comprehensive 2-DE analysis of (32)P-labelled proteins isolated from 3T3-L1 adipocytes (Hill et al. J Biol Chem 2000; 275: 24313-24320). IMPDH was identified by liquid chromatography/tandem mass spectrometry as a protein which was phosphorylated in a phosphatidylinositol (PI) 3-kinase-dependent manner upon insulin treatment. Although insulin had no significant effect on IMPDH activity, we observed translocation of IMPDH to lipid bodies following insulin treatment. Induction of lipid body formation with oleic acid promoted dramatic redistribution of IMPDH to lipid bodies, which appeared to be in contact with the endoplasmic reticulum, the site of lipid body synthesis and recycling. Inhibition of PI 3-kinase blocked insulin- and oleate-induced translocation of IMPDH and reduced oleate-induced lipid accumulation. However, we found no evidence of oleate-induced IMPDH phosphorylation, suggesting phosphorylation and translocation may not be coupled events. These data support a role for IMPDH in the dynamic regulation of lipid bodies and fatty acid metabolism and regulation of its activity by subcellular redistribution in response to extracellular factors that modify lipid metabolism.
Whitehead
Jonathan P
JP
Department of Diabetes & Obesity, Garvan Institute of Medical Research, 384 Victoria Street, Darlinghurst, Sydney, NSW 2010, Australia. jwhitehead@cder.soms.uq.edu.au
Simpson
Fiona
F
Hill
Michelle M
MM
Thomas
Elaine C
EC
Connolly
Lisa M
LM
Collart
Frank
F
Simpson
Richard J
RJ
James
David E
DE
D041721:3T3-L1 Cells; D000818:Animals; D016466:CHO Cells; D002458:Cell Fractionation; D002853:Chromatography, Liquid; D006224:Cricetinae; D003412:Cricetulus; D007168:IMP Dehydrogenase / Q000378:metabolism*; D007328:Insulin / Q000494:pharmacology*; D050356:Lipid Metabolism; D013058:Mass Spectrometry; D051379:Mice; D008856:Microscopy, Fluorescence; D019301:Oleic Acid / Q000494:pharmacology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*
D007328:Insulin; D019301:Oleic Acid; D007168:IMP Dehydrogenase; D019869:Phosphatidylinositol 3-Kinases