SUBCELLULAR-LOCALIZATION AND HORMONE SENSITIVITY OF ADIPOCYTE CYCLIC-AMP PHOSPHODIESTERASE
SUBCELLULAR-LOCALIZATION AND HORMONE SENSITIVITY OF ADIPOCYTE CYCLIC-AMP PHOSPHODIESTERASE
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DOI：
10.1042/bj2620867
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发表时间：
1989-09-15
影响因子：
4.1
通讯作者：
HOUSLAY, MD
中科院分区：
文献类型：
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作者：
ANDERSON, NG;KILGOUR, E;HOUSLAY, MD
用胰岛素和肾上腺素中的一种或两种处理完整的脂肪细胞，仅在内质网亚组分中刺激膜环AMP磷酸二酯酶活性。环GMP抑制环AMP磷酸二酯酶活性也被发现在这个馏分。定量Western印迹使用特定的多克隆抗体，提出了对同质的“致密囊泡”环AMP磷酸二酯酶从大鼠肝脏，确定了一个单一的63 kDa的物种，这是本地化的脂肪细胞内质网部分。肾上腺素刺激脂肪细胞膜环AMP磷酸二酯酶的能力显示是通过α-AMP介导的。肾上腺素受体而不是α 1-肾上腺素受体。膜环AMP磷酸二酯酶刺激胰高血糖素，但不加压素，A23187或12-O-十四烷酰佛波醇13-乙酸酯（TPA）。用氯喹或丹酰尸胺处理脂肪细胞不能影响胰岛素刺激环AMP磷酸二酯酶活性的能力。用纯化的蛋白激酶A处理分离的脂肪细胞内质网膜部分，使其环AMP磷酸二酯酶活性增加约2倍。当用纯化的蛋白激酶A和[32 P]ATP处理该组分时，标记物被掺入63 kDa的蛋白质中，该蛋白质被抗肝“致密囊泡”环AMP磷酸二酯酶的抗血清特异性免疫沉淀。
Treatment of intact adipocytes with either or both insulin and adrenaline stimulated membrane cyclic AMP phsophodiesterase activity only in the endoplasmic reticulum subfraction. The cyclic GMP-inhibited cyclic AMP phosphodiesterase activity was also found in this fraction. Quantitative Western blotting using a specific polyclonal antibody, raised against the homogenous ''dense-vesicle'' cyclic AMP phosphodiesterase from rat liver, identified a single 63 kDa species which was localized in the adipocyte endoplasmic reticulum fraction. The ability of adrenaline to stimulate adipocyte membrane cyclic AMP phosphodiesterase was shown to be mediated via .alpha.-adrenceptors and not .alpha.1-adrenoceptors. Membrane cyclic AMP phosphodiesterase was stimulated by glucagon but not by vasopressin, A23187 or 12-O-tetradecanoylphorbol 13-acetate (TPA). Treatment of adipocytes with either chloroquine or dansyl cadaverine failed to affect the ability of insulin to stimulate cyclic AMP phosphodiesterase activity. Treatment of an isolated adipocyte endoplasmic reticulum membrane fraction with purified protein kinase A increased its cyclic AMP phosphodiesterase activity some 2-fold. When this fraction was treated with purified protein kinase A and [32P]ATP, label was incorporated into a 63 kDa protein which was specifically immunoprecipitated with the antiserum against the liver ''dense-vesicle'' cyclic AMP phosphodiesterase.