Proteins routinely undergo post-translational modifications, which are crucial for their optimal function. One such post-translational modification introduces <i>N</i>-acetylglucosamine (GlcNAc) to certain serine and threonine residues.  Determining site-specifically where the GlcNAc groups are is important to understanding the role they play in the regulation and function of proteins. Our goal is to synthesize a photocleavable linker that covalently binds to proteins bearing GlcNAc and aids in their purification.  Once the proteins are purified, the linker will be removed photochemically before mass spectrometric analysis is done to determine site-specifically which serine and threonine residues bear GlcNAc groups. The photocleavable linker contains three important components: a strained alkyne, a photocleavable group, and an insoluble bead.  The focus of our initial efforts has been on synthesizing a model of the targeted linker and exploring its photochemistry.  We have found that an aminocoumarin bearing an ester undergoes a relatively clean, reliable photocleavage to afford a carboxylic acid.  Our current focus is on introducing the strained alkyne to the aminocoumarin and exploring the behavior of the product.  Future work will involve completing the synthesis by introducing the insoluble bead to the aminocoumarin/strained alkyne portion of the linker.

Towards the synthesis of a photocleavable linker for GlcNAc-ligated protein purification

249th National Meeting (2015)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.