التفاصيل البيبلوغرافية
| العنوان: |
NMR Backbone Assignment of Large Proteins by Using 13Cα-Only Triple-Resonance Experiments. |
| المؤلفون: |
Wei, Qingtao, Chen, Jiajing, Mi, Juan, Zhang, Jiahai, Ruan, Ke, Wu, Jihui |
| المصدر: |
Chemistry - A European Journal; 7/4/2016, Vol. 22 Issue 28, p9556-9564, 9p |
| مصطلحات موضوعية: |
FORCED vibration (Mechanics), PROTEIN domains, TERTIARY structure, NUCLEAR spin, MAGNETIC resonance |
| مستخلص: |
Nuclear magnetic resonance (NMR) is a powerful tool to interrogate protein structure and dynamics residue by residue. However, the prerequisite chemical-shift assignment remains a bottleneck for large proteins due to the fast relaxation and the frequency degeneracy of the 13Cα nuclei. Herein, we present a covariance NMR strategy to assign the backbone chemical shifts by using only HN(CO)CA and HNCA spectra that has a high sensitivity even for large proteins. By using the peak linear correlation coefficient (LCC), which is a sensitive probe even for tiny chemical-shift displacements, we correctly identify the fidelity of approximately 92 % cross-peaks in the covariance spectrum, which is thus a significant improvement on the approach developed by Snyder and Brüschweiler (66 %) and the use of spectral derivatives (50 %). Thus, we calculate the 4D covariance spectrum from HN(CO)CA and HNCA experiments, in which cross-peaks with LCCs above a universal threshold are considered as true correlations. This 4D covariance spectrum enables the sequential assignment of a 42 kDa maltose binding protein (MBP), in which about 95 % residues are successfully assigned with a high accuracy of 98 %. Our LCC approach, therefore, paves the way for a residue-by-residue study of the backbone structure and dynamics of large proteins. [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
