Gupta Nutan, Duggal Shweta, Jailkhani Noor, Chatterjee Samrat, Rao Kanury V S, Kumar Ajay
International Centre for Genetic Engineering and Biotechnology (ICGEB), Aruna Asif Ali Marg, New Delhi 110067, India.
Drug Discovery Research Center (DDRC), Translational Health Science and Technology Institute (THSTI), NCR Biotech Science Cluster, 3rd Milestone, Faridabad-Gurgaon Expressway, Faridabad 121001, Haryana, India.
Data Brief. 2017 May 25;13:187-191. doi: 10.1016/j.dib.2017.05.040. eCollection 2017 Aug.
Akt1 is a multi-functional protein, implicated in multiple human solid tumors. Pertaining to its key role in cell survival, Akt1 is under focus for development of targeted therapies. Functional diversity of Akt1 is a result of its interactions with other proteins; which changes with changing context. This investigation was designed to capture the dynamics of Akt1 Interactome as a function of its active state. Delineating dynamic changes in association of Akt1 with its interactors could help us comprehend how it changes as a function of inhibition of its active form. Similar information on changes in Akt1 interactome as of now is not well explored. Akt1 expressing HEK293 cells were cultured in light and heavy labeled SILAC media. Normal lysine and arginine were incorporated as light labels while for heavy labeling the isotopes were 8 and 10 Da heavier. Light labeled cells represented the indigenous state of Akt1 interactome while heavy labeled cells represented Akt1 interactome in presence of its allosteric inhibitor, MK-2206. Equal number of cells from both conditions were pooled, lysed and subjected to Affinity Purification coupled to Mass Spectroscopy (AP-MS). Additionally, SILAC labeling aided in quantitative estimation of changing association of a number of proteins which were common to the two experimental conditions, with Akt1. Data are available via ProteomeXchange with identifier PXD005976.
Akt1是一种多功能蛋白，与多种人类实体瘤有关。鉴于其在细胞存活中的关键作用，Akt1成为靶向治疗开发的重点。Akt1的功能多样性是其与其他蛋白质相互作用的结果；这种相互作用会随着环境的变化而改变。本研究旨在捕捉Akt1相互作用组的动态变化及其活性状态的函数关系。描绘Akt1与其相互作用蛋白之间关联的动态变化，有助于我们理解其如何随着活性形式的抑制而变化。目前关于Akt1相互作用组变化的类似信息尚未得到充分探索。在轻标记和重标记的稳定同位素标记氨基酸细胞培养（SILAC）培养基中培养表达Akt1的人胚肾293细胞。正常的赖氨酸和精氨酸作为轻标记掺入，而重标记的同位素则重8和10道尔顿。轻标记的细胞代表Akt1相互作用组的原始状态，而重标记的细胞代表在其变构抑制剂MK - 2206存在下的Akt1相互作用组。将两种条件下数量相等的细胞混合、裂解，并进行亲和纯化与质谱联用（AP - MS）。此外，SILAC标记有助于定量估计两种实验条件下许多与Akt1共有的蛋白质的关联变化。数据可通过蛋白质组交换获得，标识符为PXD005976。
