Nicholas Holloran Doctoral Final Exam

About the event

Presenter: Nicholas Holloran
Group: Garner
Title: Exploration of Strategies for the Synthesis of Homogeneous N-Glycoproteins

Abstract: N-glycoproteins are complex biological structures consisting of a protein and an oligosaccharide which have been linked together by a covalent bond to the side-chain of an asparagine residue. These structures have a wide array of biological functions, and the study of these molecules has been hindered by the fact that they exist naturally as complex mixtures. These mixtures can contain proteins with various numbers of glycans (macroheterogeneity) and some proteins may exhibit different glycans from others (microheterogeneity). Because of the complexity of glycoprotein samples, examination of their biological function is difficult, and it is desirable to develop a method that is capable of generating a homogeneous sample of N-glycoprotein (i.e. every protein contains the same glycans at the same positions). This work explores various methods and strategies in our attempt to synthesize homogeneous N-glycoprotein samples. It was found that a cysteine linker between the protein and the glycan could be used to facilitate the synthesis of a cysteine-bridged glycoprotein. The cysteine bridge could also be biotinylated via disulfide bond which allowed for purification of the intact cysteine-bridged glycoprotein by avidin affinity chromatography.

Zoom Information:
https://wsu.zoom.us/s/8200631218