Tags for imaging small molecules in cells
Lead Research Organisation:
University of Strathclyde
Department Name: Pure and Applied Chemistry
Abstract
A specific challenge in chemical biology is to establish efficient small molecule target engagement in cells. Within this project we will establish tags for the imaging and photo cross linking of small molecules in cells to facilitate target validation and the development of chemical probes.
Many molecular biology based approaches for biochemical analysis require cells to be lysed and individual cell components isolated, through techniques such as the polymerase chain reaction, electrophoresis and Western blotting. While these methods can give a high level of chemically specific information, they are destructive and could introduce artefacts and modifications to the sample during the process of lysing and isolating cell components. The majority of conventional imaging strategies rely on fluorescently tagging biomolecules for the detection of specific species. This approach allows species to be imaged in their native environment non-destructively but once again, there are disadvantages associated with the need for an external label to be added to the sample, which could itself interfere with the native state of the system. In addition, the dyes available for staining tend to be large therefore having the potential to heavily alter innate distribution, dynamics and metabolism.
In order to gain insight into small molecule distribution and dynamics with regards to specific biomolecules, a combination of a labelling approach with Raman imaging could be of great use. Alkyne tags have become important functional groups for labelling species for tracking using Raman spectroscopy. This use stems from the unique alkyne signal observed in the biologically 'silent' region of the Raman spectrum between approximately 1800 and 2800 cm-1. Alkyne functionality is not an inherent characteristic of native biological systems and therefore in recent years a number of studies have created biorthogonal alkyne tagged reporters in order to image specific biological species or follow the dynamics of biological processes.
We will undertake a project to evaluate alkyne tags and optimise them to deliver an enhanced Raman signal. We will then use these tags to establish molecule distribution and target engagement in a series of classes of biological target including lipids, kinases, bromodomains and nuclear receptors.
Having established optimal Raman tags we will combine this knowledge with photoaffinity labels in order to track molecules within the cell and establish target engagement.
The project will allow for an efficient and economical method for which to establish target engagement of candidate molecules in cells to facilitate target validation and the development of effective chemical probes.
Many molecular biology based approaches for biochemical analysis require cells to be lysed and individual cell components isolated, through techniques such as the polymerase chain reaction, electrophoresis and Western blotting. While these methods can give a high level of chemically specific information, they are destructive and could introduce artefacts and modifications to the sample during the process of lysing and isolating cell components. The majority of conventional imaging strategies rely on fluorescently tagging biomolecules for the detection of specific species. This approach allows species to be imaged in their native environment non-destructively but once again, there are disadvantages associated with the need for an external label to be added to the sample, which could itself interfere with the native state of the system. In addition, the dyes available for staining tend to be large therefore having the potential to heavily alter innate distribution, dynamics and metabolism.
In order to gain insight into small molecule distribution and dynamics with regards to specific biomolecules, a combination of a labelling approach with Raman imaging could be of great use. Alkyne tags have become important functional groups for labelling species for tracking using Raman spectroscopy. This use stems from the unique alkyne signal observed in the biologically 'silent' region of the Raman spectrum between approximately 1800 and 2800 cm-1. Alkyne functionality is not an inherent characteristic of native biological systems and therefore in recent years a number of studies have created biorthogonal alkyne tagged reporters in order to image specific biological species or follow the dynamics of biological processes.
We will undertake a project to evaluate alkyne tags and optimise them to deliver an enhanced Raman signal. We will then use these tags to establish molecule distribution and target engagement in a series of classes of biological target including lipids, kinases, bromodomains and nuclear receptors.
Having established optimal Raman tags we will combine this knowledge with photoaffinity labels in order to track molecules within the cell and establish target engagement.
The project will allow for an efficient and economical method for which to establish target engagement of candidate molecules in cells to facilitate target validation and the development of effective chemical probes.