Development of novel probes for glycoproteomics
Date Issued
2009
Date
2009
Author(s)
Liu, Po-Yu
Abstract
Glycosylation is an important post-translational modification. It regulates the function of cells inside and outside of different biological phenomena. However, research in the biological role of carbohydrates is very difficult. The reason lies mainly in the glycoprotein, compared to protein, carbohydrate content are usually more scarce and display a heterogeneous distribution on the cell surface. In addition, the complexity of glycosylation and no template exist, make difficult to obtain pure and enough amount of carbohydrates to do analytical study. To tackle these problems, We have designed a new probe to combine the fluorescent with biotin to amplify the signal. It has three major functions: To do click chemistry via azido group, Specificity in vivo binding to labeled carbohydrates and monitor the targeted molecule by high sensitive fluorescent. Moreover, this type of probe only show high fluorescence after click reaction. This property can reduce the background and non-specific signal. We also used the high specific binding of biotin and streptavidin to enrich the labeled glycoprotein. However, the tight binding of biotin and streptavidin make the elution very difficult. To improve that, between fluorescence and biotin group, we introduced a disulfide linkage that can be cleavage by DTT or TCEP to help the purification of labeled glycoprotein. By using this method, we have succeeded to label cell by tri-functional probe and this strategy can be used in the glycomics study and hope someday we can identify the sequence of single glycoprotein. Future experiments will focus on comparative glycomics, especially the studies of glycoprotein differences in normal cells and cancer cells. Ultimately, we hope that this new type of sugar probe can be applied to the diagnosis and treatment.
Subjects
glycoproteomics
probe
fluorescence
azido
biotin
click chemistry
SDGs
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