Novel Endogenous, Insulin-Stimulated Akt2 Protein Interaction Partners in L6 Myoblasts.
Novel Endogenous, Insulin-Stimulated Akt2 Protein Interaction Partners in L6 Myoblasts.
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DOI：
10.1371/journal.pone.0140255
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Yi Z
中科院分区：
文献类型：
--
作者：
Caruso M;Zhang X;Ma D;Yang Z;Qi Y;Yi Z
胰岛素抵抗和 2 型糖尿病的特点是胰岛素信号网络中的异常反应。磷酸肌醇依赖性丝氨酸/苏氨酸激酶 Akt2 在胰岛素信号传导和葡萄糖摄取中发挥关键作用，尤其是在骨骼肌中。蛋白质-蛋白质相互作用调节 Akt2 的功能结果，进而调节 Akt2 在葡萄糖摄取中的作用。然而，在骨骼肌细胞中只发现了很少的胰岛素反应性 Akt2 相互作用伙伴。在目前的工作中，大鼠 L6 成肌细胞（一种广泛使用的胰岛素敏感骨骼肌细胞系）被用来检查内源性、胰岛素刺激的 Akt2 蛋白相互作用伙伴。 Akt2 免疫共沉淀与 1D-SDS-PAGE 结合，并通过 HPLC-ESI-MS/MS 分析组分，以揭示 Akt2 蛋白质-蛋白质相互作用。 Pull-down 测定显示了对 Akt2 同工型的特异性；未检测到 Akt1 和 Akt3 独特肽。总共检测到 49 例患者在注射胰岛素后与 Akt2 的关联显着增加 (47) 或减少 (2) (n = 4; p<0.05)。为新型 Akt2 相互作用伙伴确定了多种途径，例如 EIF2 和泛素化途径。这些数据表明，多种新的内源蛋白可能在基础条件和胰岛素刺激条件下与 Akt2 相关，从而进一步深入了解胰岛素信号网络。数据可通过 ProteomeXchange 获得，标识符为 PXD002557。
Insulin resistance and Type 2 diabetes are marked by an aberrant response in the insulin signaling network. The phosphoinositide-dependent serine/threonine kinase, Akt2, plays a key role in insulin signaling and glucose uptake, most notably within skeletal muscle. Protein-protein interaction regulates the functional consequence of Akt2 and in turn, Akt2’s role in glucose uptake. However, only few insulin-responsive Akt2 interaction partners have been identified in skeletal muscle cells. In the present work, rat L6 myoblasts, a widely used insulin sensitive skeletal muscle cell line, were used to examine endogenous, insulin-stimulated Akt2 protein interaction partners. Akt2 co-immunoprecipitation was coupled with 1D-SDS-PAGE and fractions were analyzed by HPLC-ESI-MS/MS to reveal Akt2 protein-protein interactions. The pull-down assay displayed specificity for the Akt2 isoform; Akt1 and Akt3 unique peptides were not detected. A total of 49 were detected with a significantly increased (47) or decreased (2) association with Akt2 following insulin administration (n = 4; p<0.05). Multiple pathways were identified for the novel Akt2 interaction partners, such as the EIF2 and ubiquitination pathways. These data suggest that multiple new endogenous proteins may associate with Akt2 under basal as well as insulin-stimulated conditions, providing further insight into the insulin signaling network. Data are available via ProteomeXchange with identifier PXD002557.