Traceless, non-invasive and spatiotemporal control of protein activity in cells

无痕、无创、时空控制细胞内蛋白质活性

• 批准号: BB/P009980/1
• 负责人: Rudolf Allemann
• 金额: $ 65.01万
• 依托单位: CARDIFF UNIVERSITY
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FP009980%2F1
• 关键词: Traceless; non; invasive; spatiotemporal; control

Protein characterisation is an experimental process, where scientists build a profile of the physical, chemical and biological properties of the protein of interest. Information obtained from this protein characterisation has great importance in scientific research, particularly in areas related to health, ageing and drug development. Often, the physical and chemical properties of a protein can be characterised in a test tube isolated from its natural partners in a cell. However, these studies cannot provide a full biological characterisation because the molecular environment within a cell is much more complex and contains other protein molecules, genetic material such as DNA, in addition to carbohydrates and small molecules. It would therefore be desirable to analyse target proteins in their natural environment in a cell rather than in the test tube. However, at current there are no good methods available to introduce proteins into cells and induce their activity within a cell without modifying them chemically; such modifications often alter the properties of a protein.We now wish to create a specialised tool that allows us to introduce a protein of interest into cells and subsequently activate it with infrared light, which will not damage living cells. In the past such activation has relied on ultra-violet light which is toxic to cells. Our tool will allow researchers to introduce any protein into any cell type at any time point in an unmodified form. Moreover, integrity of the biological experiment will not be affected by the use of this tool, as IR activation is harmeless and traceless. To this end we will engineer proteins to install a specialised chemical group into proteins for activation by infrared light. Highly positively-charged molecular tags will be appended to proteins, so that their solubility in water and ability to enter cell can be enhanced. Upon light activation the tag and the chemical auxiliary will be removed and the protein will be present within the cell in its natural unmodified form.We will apply our technology to activate a specific muscle-related protein, Myf-5, which upon activation with light will induce undifferentiated cells to change their morphology and turn into muscle cells. Overall our technology has the potential to activate externally supplied biologically active proteins in diseased cells and will therefore offer opportunities for future protein therapies. It therefore has huge potential for wide ranging biomedical research and commercial applications.

蛋白质表征是一个实验过程，科学家在其中建立感兴趣蛋白质的物理，化学和生物学特性的概况。从这种蛋白质表征中获得的信息在科学研究中具有重要意义，特别是在与健康、老龄化和药物开发相关的领域。通常，蛋白质的物理和化学性质可以在试管中与细胞中的天然伴侣分离。然而，这些研究不能提供完整的生物学表征，因为细胞内的分子环境要复杂得多，除了碳水化合物和小分子外，还含有其他蛋白质分子、遗传物质如DNA。因此，希望在细胞中而不是在试管中的天然环境中分析靶蛋白。然而，目前还没有很好的方法将蛋白质引入细胞并诱导其在细胞内的活性而不对它们进行化学修饰;这种修饰通常会改变蛋白质的性质。我们现在希望创造一种专门的工具，允许我们将感兴趣的蛋白质引入细胞，随后用红外光激活它，而不会损伤活细胞。在过去，这种激活依赖于对细胞有毒的紫外线。我们的工具将允许研究人员在任何时间点以未修饰的形式将任何蛋白质引入任何细胞类型。此外，生物实验的完整性不会受到使用该工具的影响，因为IR激活是无害和无痕迹的。为此，我们将设计蛋白质，将一个专门的化学基团安装到蛋白质中，以便通过红外光激活。高正电荷的分子标签将被附加到蛋白质上，使得它们在水中的溶解度和进入细胞的能力可以增强。我们将利用我们的技术来激活一种与肌肉相关的特异性蛋白质Myf-5，这种蛋白质在光激活后将诱导未分化的细胞改变其形态并转化为肌肉细胞。总的来说，我们的技术有可能激活病变细胞中外部提供的生物活性蛋白，因此将为未来的蛋白质疗法提供机会。因此，它具有广泛的生物医学研究和商业应用的巨大潜力。

项目成果

Spatio-Temporal Photoactivation of Cytotoxic Proteins.

• DOI: 10.1002/cbic.202200115
• 发表时间: 2022-06-20
• 期刊: CHEMBIOCHEM
• 影响因子: 3.2
• 作者: Cruz-Samperio, Raquel;Mart, Robert J.;Luk, Louis Y. P.;Tsai, Yu-Hsuan;Jones, Arwyn T.;Allemann, Rudolf K.
• 通讯作者: Allemann, Rudolf K.