Related Experiment Video
Updated: Jun 10, 2026
14:44
Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR
Published on: December 16, 2013
G-matrix Fourier transform NOESY-based protocol for high-quality protein structure determination.
Yang Shen1, Hanudatta S Atreya, Gaohua Liu
1Department of Chemistry, The State University of New York at Buffalo, Buffalo, New York 14260, USA.
Journal of the American Chemical Society
|June 23, 2005
Summary
A new nuclear magnetic resonance (NMR) protocol using G-matrix Fourier transform (GFT) enables rapid, high-quality protein structure determination. This method efficiently assigns Nuclear Overhauser Effect (NOE) constraints for proteins up to 25 kDa, accelerating structural biology research.
Area of Science:
- Structural Biology
- Biophysics
- Nuclear Magnetic Resonance (NMR) Spectroscopy
Background:
- High-quality protein structure determination is crucial for understanding biological function.
- Traditional NMR methods can be time-consuming, limiting throughput.
- G-matrix Fourier transform (GFT) NMR offers potential for improved efficiency.
Purpose of the Study:
- To present a novel GFT NMR protocol for rapid, high-quality protein structure determination.
- To demonstrate the protocol's effectiveness for resonance assignment and Nuclear Overhauser Effect (NOE) analysis.
- To assess the protocol's performance on a 14 kDa protein target.
Main Methods:
- Utilized five through-bond chemical shift correlation experiments for 4D and 5D spectral information.
- Implemented a (4,3)D GFT NOESY experiment encoding multiple NOESY correlations.
- Acquired through-bond correlation and NOESY spectra in under 56 hours on a 600 MHz cryogenic probe spectrometer.
Main Results:
- Achieved high digital resolution and efficient resonance assignment.
- Successfully determined a high-quality structure for the 14 kDa protein YqfB.
- Demonstrated that NOE assignment from chemical shifts yields accurate initial structures.
- Information theoretical analysis confirmed the protocol's non-redundant constraint identification.
Conclusions:
- The presented GFT NMR protocol enables rapid data collection for robust protein structure determination.
- The method is suitable for high-throughput structure determination of proteins up to 20-25 kDa.
- This approach accelerates the process of obtaining accurate protein structures using NMR.
Related Concept Videos
01:062D NMR: Homonuclear Correlation Spectroscopy (COSY)
Homonuclear correlation spectroscopy, or COSY, is a 2-dimensional NMR technique that provides information about coupled protons. Typically, the geminal and vicinal coupling are observed. For example, consider the COSY spectrum of ethyl acetate, where its 1D proton NMR spectrum is plotted along the vertical and horizontal axes with their corresponding chemical shift scale. Three spots on the diagonal corresponding to the three peaks in the 1D proton spectrum are called diagonal peaks. The COSY...
01:15¹H NMR of Conformationally Flexible Molecules: Variable-Temperature NMR
The axial and equatorial protons in cyclohexane can be distinguished by performing a variable-temperature NMR experiment. In this process, except for one proton, the remaining eleven protons are replaced by deuterium. The deuterium substitution avoids the possible peak splitting caused by the spin-spin coupling between the adjacent protons. The remaining proton flips between the axial and equatorial positions.
01:06Nuclear Overhauser Enhancement (NOE)
Irradiation of a spin-active nucleus causes an increase or decrease in the signal intensity of neighboring nuclei that are not necessarily chemically bonded or involved in J-coupling. This phenomenon, called the nuclear Overhauser enhancement (NOE), results from through-space interactions between the nuclear spins. The NOE effect decreases with increasing internuclear distance and is generally not observed beyond 4 angstroms. In NOE, dipole-dipole interactions between neighboring spin-active...
01:25Nucleic Acid Structure
The pentose sugar in DNA is deoxyribose, while in RNA the pentose sugar is ribose. The difference between the sugars is the presence of the hydroxyl group on the ribose's second carbon and a hydrogen on the deoxyribose's second carbon. The phosphate residue attaches to the hydroxyl group of the 5′ carbon of one sugar and the hydroxyl group of the 3′ carbon of the sugar of the next nucleotide, which forms a 5′ to 3′ phosphodiester linkage.
DNA Structure
DNA has a double-helix structure. The...
DNA Structure
DNA has a double-helix structure. The...
01:162D NMR: Overview of Homonuclear Correlation Techniques
Homonuclear correlation spectroscopy (COSY) is a powerful technique used in Nuclear Magnetic Resonance (NMR) spectroscopy to study the correlations between nuclei of the same type within a molecule. It provides information about scalar couplings between adjacent nuclei, which helps determine connectivity and structural information. There are several COSY variants, each with its unique strengths and experimental parameters.
COSY90 is the standard two-dimensional (2D) COSY experiment that...
COSY90 is the standard two-dimensional (2D) COSY experiment that...
01:182D NMR: Overview of Heteronuclear Correlation Techniques
Heteronuclear correlation spectroscopy is an analytical technique that investigates the coupling between different types of nuclei, often a proton and an X-nucleus, such as carbon-13 or nitrogen-15. This method is commonly used in nuclear magnetic resonance (NMR) spectroscopy to gain insights into complex chemical compounds' structural and compositional aspects. A typical heteronuclear correlation spectrum displays X-nucleus chemical shifts on one axis and a proton spectrum on the other axis.