Department of Cellular and Molecular Pharmacology, Institute of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan; Department of Oral Surgery, Institute of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.
Department of Cellular and Molecular Pharmacology, Institute of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.
PLoS One. 2014 Jun 19;9(6):e100559. doi: 10.1371/journal.pone.0100559. eCollection 2014.
Phosphorylation of hormone-sensitive lipase (HSL) and perilipin by protein kinase A (PKA) promotes the hydrolysis of lipids in adipocytes. Although activation of lipolysis by PKA has been well studied, inactivation via protein phosphatases is poorly understood. Here, we investigated whether phospholipase C-related catalytically inactive protein (PRIP), a binding partner for protein phosphatase 1 and protein phosphatase 2A (PP2A), is involved in lipolysis by regulating phosphatase activity. PRIP knockout (PRIP-KO) mice displayed reduced body-fat mass as compared with wild-type mice fed with standard chow ad libitum. Most other organs appeared normal, suggesting that mutant mice had aberrant fat metabolism in adipocytes. HSL in PRIP-KO adipose tissue was highly phosphorylated compared to that in wild-type mice. Starvation of wild-type mice or stimulation of adipose tissue explants with the catabolic hormone, adrenaline, translocated both PRIP and PP2A from the cytosol to lipid droplets, but the translocation of PP2A was significantly reduced in PRIP-KO adipocytes. Consistently, the phosphatase activity associated with lipid droplet fraction in PRIP-KO adipocytes was significantly reduced and was independent of adrenaline stimulation. Lipolysis activity, as assessed by measurement of non-esterified fatty acids and glycerol, was higher in PRIP-KO adipocytes. When wild-type adipocytes were treated with a phosphatase inhibitor, they showed a high lipolysis activity at the similar level to PRIP-KO adipocytes. Collectively, these results suggest that PRIP promotes the translocation of phosphatases to lipid droplets to trigger the dephosphorylation of HSL and perilipin A, thus reducing PKA-mediated lipolysis.
蛋白激酶 A(PKA)对激素敏感脂肪酶(HSL)和脂滴包被蛋白( perilipin)的磷酸化作用促进了脂肪细胞中脂质的水解。尽管 PKA 对脂肪分解的激活作用已得到充分研究,但通过蛋白磷酸酶失活的机制却知之甚少。在这里,我们研究了磷脂酶 C 相关无催化活性蛋白(PRIP)是否通过调节磷酸酶活性参与脂肪分解,因为它是蛋白磷酸酶 1 和蛋白磷酸酶 2A(PP2A)的结合伴侣。与自由进食标准饲料的野生型小鼠相比,PRIP 敲除(PRIP-KO)小鼠的体脂质量减少。大多数其他器官似乎正常,这表明突变小鼠的脂肪代谢异常。与野生型小鼠相比,PRIP-KO 脂肪组织中的 HSL 高度磷酸化。饥饿或用分解代谢激素肾上腺素刺激野生型小鼠的脂肪组织外植体,将 PRIP 和 PP2A 从细胞质转位到脂滴,但在 PRIP-KO 脂肪细胞中 PP2A 的转位明显减少。一致地,PRIP-KO 脂肪细胞中与脂滴部分相关的磷酸酶活性显著降低,并且与肾上腺素刺激无关。通过测量非酯化脂肪酸和甘油来评估脂肪分解活性,PRIP-KO 脂肪细胞中的活性更高。当用磷酸酶抑制剂处理野生型脂肪细胞时,它们显示出与 PRIP-KO 脂肪细胞相似的高水平脂肪分解活性。总之,这些结果表明 PRIP 促进了磷酸酶向脂滴的转位,以触发 HSL 和 perilipin A 的去磷酸化,从而减少 PKA 介导的脂肪分解。