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

点击翻译按钮获取中文摘要

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.