Denaturant-induced Expansion and Compaction of a Multi-domain Protein: IgG
| Autor: | Pramit K. Chowdhury, Julie M. Glasscock, Lin Guo, Feng Gai |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Protein Denaturation
Protein Folding Time Factors Hydrodynamic radius Diffusion Fluorescence correlation spectroscopy Article Immunoglobulin Fab Fragments chemistry.chemical_compound Structural Biology Animals Guanidine Molecular Biology biology Goats Protein Structure Tertiary Solvent Crystallography chemistry Immunoglobulin G Radius of gyration biology.protein Protein folding Protein A |
| Zdroj: | Journal of Molecular Biology. 384:1029-1036 |
| ISSN: | 0022-2836 |
| DOI: | 10.1016/j.jmb.2008.03.006 |
| Popis: | It is generally believed that unfolded or denatured proteins show random-coil statistics and hence their radius of gyration simply scales with solvent quality (or concentration of denaturant). Indeed, nearly all proteins studied thus far have been shown to undergo a gradual and continuous expansion with increasing concentration of denaturant. Here, we use fluorescence correlation spectroscopy (FCS) to show that while protein A, a multi-domain and predominantly helical protein, expands gradually and continuously with increasing concentration of guanidine hydrochloride (GdnHCl), the F(ab')2 fragment of goat anti-rabbit antibody IgG, a multi-subunit all beta-sheet protein does not show such continuous expansion behavior. Instead, it first expands and then contracts with increasing concentration of GdnHCl. Even more striking is the fact that the hydrodynamic radius of the most expanded F(ab')2 ensemble, observed at 3-4 M GdnHCl, is approximately 3.6 times that of the native protein. Further FCS measurements involving urea and NaCl show that the unusually expanded F(ab')2 conformations might be due to electrostatic repulsions. Taken together, these results suggest that specific interactions need to be considered while assessing the conformational and statistical properties of unfolded proteins, particularly under conditions of low solvent quality. |
| Databáze: | OpenAIRE |
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