Hormonal regulation of phosphodiesterase 3B in adipocytes

Detta är en avhandling från Lund University, Medical Faculty, Department of Experimental Medical Science

Sammanfattning: Insulin induced activation of the cAMP degrading enzyme phosphodiesterase (PDE) 3B in adipocytes is important for insulin mediated inhibition of lipolysis. The aim of this thesis was to contribute with knowledge on the acute activation of PDE3B in response to insulin and catecholamines in adipocytes. The activation of PDE3B was investigated from three different perspectives namely; phosphorylation of PDE3B, localization of PDE3B and PDE3B interactions with other proteins. As models to study activation of PDE3B we used primary rat and mouse adipocytes, 3T3-L1 adipocytes and caveolin-1 knock out mouse adipocytes. The study on PDE3B phosphorylation shows that PDE3B is multisite phosphorylated in response to both insulin and catecholamines. We were able to identify six phosphorylation sites on PDE3B namely; 273, S296, S421, S424/5, S474 and S536. From the studies on PDE3B localization we conclude that PDE3B is localized to caveolae and endoplasmic reticulum (ER) in adipocytes and that caveolae localized PDE3B is specifically activated by catecholamines while PDE3B in ER is specifically activated by insulin. Finally, the studies on PDE3B interactions demonstrate that insulin and catecholamines stimulate the recruitment of PDE3B into large macromolecular complexes in adipocytes. The recruitment is dependent on caveolin-1 especially in response to insulin and the constituents of the PDE3B macromolecular complexes are dependent on the stimuli used. After insulin stimulation insulin signaling molecules such as protein kinase B (PKB) are associated with PDE3B in the large complex while after catecholamine stimulation adrenergic signaling molecules such as protein kinase A (PKA) are associated with PDE3B.