Neuroprotection Research Laboratory, Departments of Neurology and Radiology, Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School, Charlestown, MA, USA.
Neuroscience. 2012 Jan 3;200:99-105. doi: 10.1016/j.neuroscience.2011.10.046. Epub 2011 Nov 3.
Neuroglobin (Ngb) is a globin protein that is highly and specifically expressed in brain neurons. A large volume of evidence has proven that Ngb is a neuroprotective molecule against hypoxic/ischemic brain injury and other related neurological disorder; however, the underlying mechanisms remain poorly understood. Aiming to provide more clues in understanding the molecular mechanisms of Ngb's neuroprotection, we performed yeast two-hybrid screening to search for proteins that interact with Ngb. From a mouse brain cDNA library, we found totally 36 proteins that potentially interact with Ngb, and 10 of them were each identified in multiple positive clones. The shared sequences within these multiple clones are more likely to be Ngb-interacting domains. In primary cultured mouse cortical neurons, immuno-precipitation was performed to confirm the interactions of selected proteins with Ngb. The discovered Ngb-interacting proteins in this study include those involved in energy metabolism, mitochondria function, and signaling pathways for cell survival and proliferation. Our findings provide molecular targets for investigating protein interaction-based biological functions and neuroprotective mechanisms of Ngb.
神经球蛋白（Ngb）是一种球蛋白蛋白，在脑神经元中高度特异性表达。大量证据证明，Ngb 是一种对缺氧/缺血性脑损伤和其他相关神经疾病具有神经保护作用的分子；然而，其潜在的机制仍知之甚少。为了提供更多的线索来理解 Ngb 神经保护的分子机制，我们进行了酵母双杂交筛选，以寻找与 Ngb 相互作用的蛋白质。从小鼠脑 cDNA 文库中，我们发现了总共 36 种可能与 Ngb 相互作用的蛋白质，其中 10 种在多个阳性克隆中被鉴定出来。这些多个克隆中的共享序列更有可能是 Ngb 相互作用的结构域。在原代培养的小鼠皮质神经元中，进行免疫沉淀以确认所选蛋白质与 Ngb 的相互作用。本研究中发现的与 Ngb 相互作用的蛋白质包括参与能量代谢、线粒体功能以及细胞存活和增殖信号通路的蛋白质。我们的发现为研究基于蛋白质相互作用的生物学功能和 Ngb 的神经保护机制提供了分子靶标。
