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Diabetes 54:3349-3357, 2005
© 2005 by the American Diabetes Association, Inc.

Expression of Inducible 6-Phosphofructo-2-Kinase/Fructose-2,6-Bisphosphatase/PFKFB3 Isoforms in Adipocytes and Their Potential Role in Glycolytic Regulation

Toshiya Atsumi1, Taro Nishio1, Hirokatsu Niwa1, Jun Takeuchi1, Hidenori Bando1, Chikara Shimizu1, Narihito Yoshioka1, Richard Bucala2, and Takao Koike1

1 Department of Medicine II, Graduate School of Medicine, Hokkaido University, Sapporo, Japan
2 Department of Medicine and Pathology, School of Medicine, Yale University, New Haven, Connecticut

6-Phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK-2/FBPase) catalyzes the synthesis and degradation of fructose 2,6-bisphosphate (F2,6BP), which is a powerful activator of 6-phosphofructo-1-kinase, the rate-limiting enzyme of glycolysis. Four genes encode PFK-2/FBPase (PFKFB1–4), and an inducible isoform (iPFK-2/PFKFB3) has been found to mediate F2,6BP production in proliferating cells. We have investigated the role of iPFK-2/PFKFB3 and related isoforms in the regulation of glycolysis in adipocytes. Human visceral fat cells express PFKFB3 mRNA, and three alternatively spliced isoforms of iPFK-2/PFKFB3 are expressed in the epididymal fat pad of the mouse. Forced expression of the iPFK-2/PFKFB3 in COS-7 cells resulted in increased glucose uptake and cellular F2,6BP content. Prolonged insulin treatment of 3T3-L1 adipocytes led to reduced PFKFB3 mRNA expression, and epididymal fat pads from db/db mice also showed decreased expression of PFKFB3 mRNA. Finally, anti–phospho-iPFK-2(Ser461) Western blotting revealed strong reactivity in insulin-treated 3T3-L1 adipocyte, suggesting that insulin induces the phosphorylation of PFKFB3 protein. These data expand the role of these structurally unique iPFK-2/PFKFB3 isoforms in the metabolic regulation of adipocytes.

Address correspondence and reprint requests to Toshiya Atsumi, MD, PhD, Department of Medicine II, Hokkaido University Graduate School of Medicine, kita 15, Nishi 7, kita-ku, Sapporo 060-8638, Japan. E-mail: tatsumi{at}med.hokudai.ac.jp

Abbreviations: DMEM, Dulbecco’s modified Eagle’s medium; F2,6BP, fructose 2,6-bisphosphate; FBS, fetal bovine serum; IBMX, 3-isobutyl-1-methylxanthine; IRS, insulin receptor substrate; PFK-1, phosphofructokinase-1; PFK-2/FBPase, 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase; PPAR, peroxisome proliferator–activated receptor; UTR, untranslated region


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