The overall objectives of this research project are to ascertain the principles that determine binding affinity and specificity in the recognition of the tetrasaccharide sialyl-Lewis x by the E-selectin receptor and to generate molecules that inhibit this interaction. Sialyl-Lewis x is found on the surface of leukocytes, and the E-selectin receptor is expressed on endothelial cells. Mounting evidence suggests that the formation of a protein-complex between these molecules, mediates call adhesion between leukocytes and the vascular endothelium. This intercellular interaction is an early event in an inflammatory or immune response. Inhibitors of this adhesive interaction could be used to develop new strategies for treatment of inflammatory conditions such as rheumatoid arthritis, multiple sclerosis, and reperfusion injury. These cell adhesion processes represent fundamentally new therapeutic targets. The generation of molecules to interfere with carbohydrate function offers a new frontier in inhibitor design. The proposed approach involves combining concepts and synthetic methods from organic chemistry with techniques from biochemistry and molecular and structural biology to generate the component molecules and to study the structure-function relationships in this system.
The specific aims of this research are as follows: (1) To use organic synthesis to generate the naturally-occurring carbohydrate sialyl-Lewis x and to investigate the folding properties of this carbohydrate by multi- dimensional NMR spectroscopy, (2) To produce the region of the E-selectin receptor that is responsible for carbohydrate-binding using a fusion expression system, which will allow for the generation of the free protein for NMR studies and immobilized protein for carbohydrate-binding studies, (3) To study the conformation of the oligosaccharide bound to the carbohydrate recognition domain of the E-selectin protein by multi- dimensional NMR spectroscopy, (4) To synthesize polymers containing carbohydrate residues in defined locations as potential multivalent inhibitors of cell adhesion. The information that is gained from the studies of the complex will be incorporated into the design of anti-adhesive compounds. The proposed research will generate insight into the molecular recognition and biological role for oligosaccharides in cell adhesion. It is anticipated that these studies will lead to the development of new methodologies for the modulation of cell-cell and carbohydrate-receptor interactions.
| Xu, Yongmei; Chandarajoti, Kasemsiri; Zhang, Xing et al. (2017) Synthetic oligosaccharides can replace animal-sourced low-molecular weight heparins. Sci Transl Med 9: |
| Hudson, Kieran L; Bartlett, Gail J; Diehl, Roger C et al. (2015) Carbohydrate-Aromatic Interactions in Proteins. J Am Chem Soc 137:15152-60 |
| Bennett, Nitasha R; Zwick, Daniel B; Courtney, Adam H et al. (2015) Multivalent Antigens for Promoting B and T Cell Activation. ACS Chem Biol 10:1817-24 |
| Wrighton, Paul J; Kiessling, Laura L (2015) Forces of Change: Mechanics Underlying Formation of Functional 3D Organ Buds. Cell Stem Cell 16:453-4 |
| Wrighton, Paul J; Klim, Joseph R; Hernandez, Brandon A et al. (2014) Signals from the surface modulate differentiation of human pluripotent stem cells through glycosaminoglycans and integrins. Proc Natl Acad Sci U S A 111:18126-31 |
| Sheridan, Rachael T C; Hudon, Jonathan; Hank, Jacquelyn A et al. (2014) Rhamnose glycoconjugates for the recruitment of endogenous anti-carbohydrate antibodies to tumor cells. Chembiochem 15:1393-8 |
| Courtney, Adam H; Bennett, Nitasha R; Zwick, Daniel B et al. (2014) Synthetic antigens reveal dynamics of BCR endocytosis during inhibitory signaling. ACS Chem Biol 9:202-10 |
| Musah, Samira; Wrighton, Paul J; Zaltsman, Yefim et al. (2014) Substratum-induced differentiation of human pluripotent stem cells reveals the coactivator YAP is a potent regulator of neuronal specification. Proc Natl Acad Sci U S A 111:13805-10 |
| Fishman, Joshua M; Kiessling, Laura L (2013) Synthesis of functionalizable and degradable polymers by ring-opening metathesis polymerization. Angew Chem Int Ed Engl 52:5061-4 |
| Kiessling, Laura L; Grim, Joseph C (2013) Glycopolymer probes of signal transduction. Chem Soc Rev 42:4476-91 |
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