Quantifying microsecond exchange in large protein complexes with accelerated relaxation dispersion experiments in the solid state
| Autor: | Carl Öster, Simone Kosol, Józef R. Lewandowski |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Protein Conformation
alpha-Helical 0301 basic medicine Materials science Analytical chemistry lcsh:Medicine Solid-state NMR Article Slow motion Motion 03 medical and health sciences Paramagnetism 0302 clinical medicine Biophysical chemistry Humans QD Solubility lcsh:Science Nuclear Magnetic Resonance Biomolecular Multidisciplinary Spectrometer lcsh:R Relaxation (NMR) Proteins QP Microsecond 030104 developmental biology Solid-state nuclear magnetic resonance Immunoglobulin G lcsh:Q Protein Conformation beta-Strand Dispersion (chemistry) 030217 neurology & neurosurgery |
| Zdroj: | Scientific Reports Scientific Reports, Vol 9, Iss 1, Pp 1-10 (2019) |
| ISSN: | 2045-2322 |
| Popis: | Solid state NMR is a powerful method to obtain information on the structure and dynamics of protein complexes that, due to solubility and size limitations, cannot be achieved by other methods. Here, we present an approach that allows the quantification of microsecond conformational exchange in large protein complexes by using a paramagnetic agent to accelerate 15N R1ρ relaxation dispersion measurements and overcome sensitivity limitations. The method is validated on crystalline GB1 and then applied to a >300 kDa precipitated complex of GB1 with full length human immunoglobulin G (IgG). The addition of a paramagnetic agent increased the signal to noise ratio per time unit by a factor of 5, which allowed full relaxation dispersion curves to be recorded on a sample containing less than 50 μg of labelled material in 5 and 10 days on 850 and 700 MHz spectrometers, respectively. We discover a similar exchange process across the β-sheet in GB1 in crystals and in complex with IgG. However, the slow motion observed for a number of residues in the α-helix of crystalline GB1 is not detected in the complex. |
| Databáze: | OpenAIRE |
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