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¹H NMR: Complex Splitting

¹H NMR: Complex Splitting

A proton M that is coupled to a proton X results in doublet signals for M. However, NMR-active nuclei can be simultaneously coupled to more than one nonequivalent nucleus. When M is coupled to a second proton A, such as in styrene oxide, each peak in the doublet is split into another doublet.
Splitting diagrams or splitting tree diagrams are routinely used to depict such complex couplings. While drawing splitting diagrams, the splitting with the larger coupling constant is usually applied...

Interpreting ¹H NMR Signal Splitting: The (n + 1) Rule

Interpreting ¹H NMR Signal Splitting: The (n + 1) Rule

In the AX proton spin system, proton A can sense the two spin states of a coupled proton X, resulting in a doublet NMR signal with two peaks of equal (1:1) intensity. When proton A is coupled to two equivalent protons (AX2 spin system), the spin states of each X can be aligned with or against the external field, creating three possible scenarios. This results in a 1:2:1 triplet signal, where the central peak corresponds to the chemical shift of A and is twice as large or intense as the...

¹H NMR: Interpreting Distorted and Overlapping Signals

¹H NMR: Interpreting Distorted and Overlapping Signals

Spin systems where the difference in chemical shifts of the coupled nuclei is greater than ten times J are called first-order spin systems. These nuclei are weakly coupled, and their chemical shifts and coupling constant can generally be estimated from the well-separated signals in the spectrum.
As Δν decreases and the signals move closer, the doublets appear increasingly distorted. The intensities of the inner lines increase at the cost of those of the outer lines as the signals are...

NMR Spectrometers: Overview

NMR Spectrometers: Overview

NMR spectrometers consist of a strong magnet, a radiofrequency transmitter, and a detector attached to a computer console for recording spectra of samples containing NMR-active nuclei. In first-generation NMR instruments called continuous-wave spectrometers, the resonance frequencies of the nuclei are determined by frequency-sweep or field-sweep methods. The magnetic field strength is fixed and the rf signal is swept in the former, while the radiofrequency signal is fixed and the magnetic field...

¹H NMR of Conformationally Flexible Molecules: Variable-Temperature NMR

¹H 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.

¹H NMR of Conformationally Flexible Molecules: Temporal Resolution

¹H NMR of Conformationally Flexible Molecules: Temporal Resolution

At room temperature, the chair conformer of cyclohexane undergoes rapid ring flipping between two equivalent chair conformers at a rate of approximately 105 times per second. These two chair conformers are in equilibrium. The rapid ring flipping results in the interconversion of the axial proton to an equatorial proton and an equatorial to the axial proton. Such interconversions are too rapid and cannot be detected on the NMR timescale. Hence, the NMR spectrometer cannot distinguish between the...

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Related Experiment Video

Updated: Feb 18, 2026

Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR

Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR

Published on: December 16, 2013

Protein Structure Elucidation from NMR Data with the Program Xplor-NIH.

Guillermo A Bermejo¹, Charles D Schwieters²

¹Center for Information Technology, National Institutes of Health, 12 South Drive, MSC 5624, Bethesda, MD, 20892, USA. guillermo.bermejo@nih.gov.

Methods in Molecular Biology (Clifton, N.J.)

|November 20, 2017

Summary

This summary is machine-generated.

This study details using Xplor-NIH software for de novo biomolecular structure determination. It demonstrates calculating the GB1 protein structure using Nuclear Magnetic Resonance data and advanced algorithms.

Keywords:

NMR Protein structure calculation Xplor-NIH script

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Area of Science:

Biomolecular NMR spectroscopy
Structural biology
Computational chemistry

Background:

Xplor-NIH is a widely used software package for determining biomolecular structures.
Nuclear Magnetic Resonance (NMR) data, including Nuclear Overhauser Effects (NOEs), scalar couplings, and Residual Dipolar Couplings (RDCs), are crucial for structure determination.
The B1 domain of streptococcal protein G (GB1) is a common model system in structural biology.

Purpose of the Study:

To illustrate the application of Xplor-NIH for de novo structure determination.
To provide a detailed example using the GB1 protein domain.
To showcase the integration of various NMR data types within the structure calculation process.

Main Methods:

De novo structure determination using Xplor-NIH software.
Utilizing Nuclear Overhauser Effect (NOE) derived distance restraints.
Incorporating torsion angle restraints from scalar couplings.
Applying bond-vector orientation restraints from Residual Dipolar Couplings (RDCs).

Main Results:

Successful de novo structure determination of the GB1 protein domain.
Demonstration of Xplor-NIH's capabilities with integrated NMR data.
A complete structure calculation script is provided as a template.

Conclusions:

Xplor-NIH is a powerful tool for biomolecular structure determination from NMR data.
The presented methodology provides a robust framework for calculating protein structures.
The detailed script serves as a valuable resource for researchers applying Xplor-NIH to other systems.