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1. Introduction
2. Protein-glycan interactions
3. Reasons for new methodology
4. Chemokine CCL5 interactions with GAGs
5. Crystal structure of a CCL5 dimer
6. NMR provides ligand binding information
7. STD-NOESY of 444 complex
8. Building models with a ligand
9. NMR works well for studies of smaller systems
10. Residual dipolar coupling (RDC)
11. WT CCL5 at low pH
12. RDC data for E66S and WT CCL5
13. Multiple models for the tetramer structure
14. SAXS data yield additional shape restrictions
15. Fixing dimer orientations
16. Cross-saturation confirms interface residues
17. MS hydroxyl radical footprinting
18. Radical footprinting & NMR cross-saturation
19. Tetramer model extends to higher oligomers
20. Modeling in longer GAG oligomers
21. Glycans on a glycoprotein
22. Why do we care about IgG glycosylation?
23. How do glycans modulate receptor interactions?
24. Glycans on native IgG-Fc are very heterogeneous
25. Investigating differential branch accessibility
26. Preparing IgG with homogeneous 13C glycans (1)
27. Production of 13C labeled IgG-Fc fragment
28. 1-3 branch is mobile and 1-6 branch is restrained
29. Relaxation dispersion probes slow exchange
30. Relaxation dispersion and chemical shift data
31. Model for galactose terminated Fc glycans
32. Locating glycans more precisely
33. Predicted distances based on 1IGT structure
34. Gal PREs in the Dy+++ : Z-L2LBT: Fc complex
35. Sialylated glycans and rheumatoid arthritis
36. Sialic acid can be produced in a labeled form
37. Preparing IgG with homogeneous 13C glycans (2)
38. 13C-Gal labeled Fc + & - sialic acid
39. Sialic acid addition & observation of NMR spectra
40. CPMG R2 relaxation can be measured
41. Di-sialylation & shift to glycan-exposed state
42. Summary
43. Acknowledgments

Topics Covered

• NMR studies
• Protein structure
• Protein-protein Interactions
• Building models with a ligand
• Chemokine CCL5 interactions with GAGs
• STD-NOESY
• Residual dipolar coupling (RDC)
• SAXS data
• Cross-saturation
• Radical footprinting
• Glycans on a glycoprotein
• - Preparing IgG with homogeneous 13C glycans
• Relaxation dispersion
• Galactose terminated Fc glycans Locating glycans more precisely
• Sialylated glycans
• CPMG R2 relaxation
• Di-sialylation

Links

Series:

• Glycobiology

Categories:

• Biochemistry
• Cell Biology

Talk Citation

Publication History

• Published on November 27, 2012

Financial Disclosures

• Prof. James Prestegard has not informed HSTalks of any commercial/financial relationship that it is appropriate to disclose.