Epitope mapping of sialyl Lewis(x) bound to E-selectin using saturation transfer difference NMR experiments
| Autor: | Beat Ernst, Meike Rinnbauer, John L. Magnani, Andrew J. Benie, Thomas Peters, Bea Wagner |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Models
Molecular Magnetic Resonance Spectroscopy Stereochemistry Molecular Sequence Data Binding pocket Oligosaccharides Peptide Nuclear Overhauser effect Biochemistry chemistry.chemical_compound Peptide Library Inhibitory peptide E-selectin Carbohydrate Conformation Sialyl Lewis X Antigen chemistry.chemical_classification biology Molecular Structure Epitope mapping Sialyl-Lewis X chemistry Carbohydrate Sequence Saturation transfer biology.protein E-Selectin Epitope Mapping Protein Binding |
| Zdroj: | Glycobiology. 13(6) |
| ISSN: | 0959-6658 |
| Popis: | A complex between sialyl Lewisx (alpha-D-Neu5Ac-[2--3]- beta-D-Gal-[1--4]-[alpha-L-Fuc-(1--3)]-beta-D-GlcNAc-O-[CH2]8 COOMe) and E-selectin was studied using saturation transfer difference (STD) nuclear magnetic resonance (NMR) experiments. These experiments allow the identification of the binding epitope of a ligand at atomic resolution. A semiquantitative analysis of STD total correlation spectroscopy spectra provides clear evidence that the galactose residue receives the largest saturation transfer. The protons H4 and H6 of the galactose residue are in especially close contact to the amino acids of the E-selectin binding pocket. The fucose residue also receives a significant saturation transfer. The GlcNAc and Neu5Ac residues, with the exception of H3 and H3' of Neu5Ac, were found to interact weakly with the protein surface. These findings are in excellent agreement with a recently published X-ray structure and with the earlier findings from syntheses and activity assays. To further characterize the binding pocket of E-selectin, an inhibitory peptide, Ac-TWDQLWDLMK-CONH2, was synthesized and the binding to E-selectin studied utilizing transfer nuclear Overhauser effect spectroscopy (trNOESY) experiments. Finally, competitive trNOESY experiments were performed, showing that the synthetic peptide is a competitive inhibitor of sialyl Lewisx. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|