Identification of the Insulin-Regulated Interaction of Phosphodiesterase 3B With 14-3-3 Protein

نویسندگان

  • Hiroshi Onuma
  • Haruhiko Osawa
  • Kazuya Yamada
  • Takahiro Ogura
  • Fumiko Tanabe
  • Daryl K. Granner
  • Hideichi Makino
چکیده

Phosphodiesterase (PDE)-3B, a major PDE isoform in adipocytes, plays a pivotal role in the antilipolytic action of insulin. Insulin-induced phosphorylation and activation of PDE3B is phosphatidylinositol 3-kinase (PI3-K) and Akt dependent, but the precise mechanism of PDE3B activation is not fully understood. We have identified 14-3-3 , a critical scaffolding molecule in signal transduction, as a protein that interacts with PDE3B using the yeast two-hybrid system. The interaction between PDE3B and 14-3-3 was then confirmed in vitro. The glutathione S-transferase (GST)-tagged 143-3 interacts with endogenous PDE3B of rat adipocytes, and this interaction is enhanced when adipocytes are treated with insulin. Coimmunoprecipitation experiments reveal that endogenous PDE3B also associates with endogenous 14-3-3 in rat adipocytes, and this interaction is enhanced by insulin. Two different PI3-K inhibitors, wortmannin and Ly294002, block this induction, suggesting that PI3-K is required. Synthetic 15 amino acid peptides of rat PDE3B containing phosphorylated Ser-279 or -302 inhibit this interaction, indicating that the insulin-regulated phosphorylation of these serine residues is involved. Because insulin receptor substrate-1 also associates with 14-3-3, the dimeric 14-3-3 could function as a scaffolding protein in the activation of PDE3B by insulin. Diabetes 51:3362–3367, 2002

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تاریخ انتشار 2002