Accelerating NMR-Based Structural Studies of Proteins by Combining Amino Acid Selective Unlabeling and Fast NMR Methods

Abstract

Abstract:In recent years, there has been a growing interest in fast acquisition and analysis of nuclear magnetic resonance (NMR) spectroscopy data for high throughput protein structure determination. Towards this end, rapid data collection techniques and methods to simplify the NMR spectrum such as amino acid selective unlabeling have been proposed recently. Combining these two approaches can speed up further the structure determination process. Based on this idea, we present three new two-dimensional (2D) NMR experiments, which together provide 15 N, 1 HN, 13 C α , 13 C β , 13 C chemical shifts for amino acid residues which are immediate C-terminal neighbors (i + 1) of residues that are selectively unlabeled. These experiments have high sensitivity and can be acquired rapidly using the methodology of G-matrix Fourier transform (GFT) NMR spectroscopy combined with non-uniform sampling (NUS). This is a first study involving the application of fast NMR methods to proteins samples prepared using a specific labeling scheme. Taken together, this opens up new avenues to using the method of selective unlabeling for rapid resonance assignment of proteins.

Nuclear Magnetic Resonance Spectroscopy

“…Involves the application of fast NMR methods, G-matrix Fourier transform (GFT) and non-uniform sampling (NUS), to proteins samples prepared using a specific labeling scheme. The spectral simplification obtained combined with rapid data collection can help in reducing the time required for data analysis [8]. (9) Supporting Protein Stability.…”