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Related Concept Videos

¹H NMR of Conformationally Flexible Molecules: Temporal Resolution00:52

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...
¹H NMR of Conformationally Flexible Molecules: Variable-Temperature NMR01:15

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: Complex Splitting01:13

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 first.
¹H NMR: Interpreting Distorted and Overlapping Signals01:02

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 slanted or...
High-Resolution Mass Spectrometry (HRMS)01:15

High-Resolution Mass Spectrometry (HRMS)

The resolution of a mass spectrometer depends on the efficiency of separating ions with different ion masses. The mass of an atom is approximated to the sum of the masses of protons and neutrons inside, considering the masses of protons and neutrons as equal. However, the masses of the proton (1.6726 × 10−24 g) and neutron (1.6749 × 10−24 g) are not truly equal. There is a minor error in the expression of atomic masses relative to the simplest atom of hydrogen. For example, the mass of helium...
Chemical Shift: Internal References and Solvent Effects01:17

Chemical Shift: Internal References and Solvent Effects

In an NMR sample, precise measurement of the absolute absorption frequencies of nuclei is difficult. A standard internal reference compound is added, and the frequency difference between the reference signal and sample signals is measured.
The internal reference compound generally used in NMR spectroscopy is tetramethylsilane (TMS). TMS is preferred because it is chemically inert, soluble in NMR solvents, and easily removable. Also, the highly shielded methyl protons in TMS yield an intense...

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Atomic Scale Structural Studies of Macromolecular Assemblies by Solid-state Nuclear Magnetic Resonance Spectroscopy
14:55

Atomic Scale Structural Studies of Macromolecular Assemblies by Solid-state Nuclear Magnetic Resonance Spectroscopy

Published on: September 17, 2017

Solution NMR of large molecules and assemblies.

Mark P Foster1, Craig A McElroy, Carlos D Amero

  • 1Department of Biochemistry, The Ohio State University, 484 West 12th Avenue, Columbus, Ohio 43210, USA. foster.281@osu.edu

Biochemistry
|January 11, 2007
PubMed
Summary

Solution NMR spectroscopy now enables detailed analysis of larger biological macromolecules and complexes. Recent advances in NMR techniques and isotopic labeling have significantly expanded the scope of NMR-tractable targets for biochemical studies.

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Last Updated: Jul 17, 2026

Atomic Scale Structural Studies of Macromolecular Assemblies by Solid-state Nuclear Magnetic Resonance Spectroscopy
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Area of Science:

  • Biophysical Chemistry
  • Structural Biology
  • Molecular Biophysics

Background:

  • Solution Nuclear Magnetic Resonance (NMR) spectroscopy is a key technique for studying biological macromolecules.
  • Multidimensional NMR and isotopic labeling (15N, 13C) have enabled detailed analysis of smaller macromolecules (< 25 kDa).

Purpose of the Study:

  • To describe methodological advances enabling NMR spectroscopy of large macromolecules and their complexes.
  • To provide a perspective on the broad applications of NMR in biochemistry.

Main Methods:

  • Utilizing multidimensional (2D-4D) NMR spectroscopy.
  • Employing uniform isotopic labeling of proteins and RNA with 15N and 13C.
  • Highlighting recent advancements in NMR and labeling strategies.

Main Results:

  • NMR is now applicable to a broader range of larger biological macromolecules and complexes.
  • Methodological improvements have expanded NMR-tractable targets by at least an order of magnitude.
  • NMR applications in biochemistry have been significantly broadened.

Conclusions:

  • Recent advances have greatly enhanced the capability of solution NMR spectroscopy for studying large biomolecules.
  • NMR spectroscopy is a versatile and powerful tool for investigating macromolecular structure, dynamics, and interactions in biochemistry.