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Related Concept Videos

¹³C NMR: Distortionless Enhancement by Polarization Transfer (DEPT)01:20

¹³C NMR: Distortionless Enhancement by Polarization Transfer (DEPT)

When proton-coupled carbon-13 spectra are simplified by a broadband proton decoupling technique, structural information about the coupled protons is lost. Distortionless enhancement by polarization transfer (DEPT) is a technique that provides information on the number of hydrogens attached to each carbon in a molecule. While the DEPT experiment utilizes complex pulse sequences, the pulse delay and flip angle are specifically manipulated. The resulting signals have different phases depending on...
IR Spectrum Peak Splitting: Symmetric vs Asymmetric Vibrations01:08

IR Spectrum Peak Splitting: Symmetric vs Asymmetric Vibrations

Identical bonds within a polyatomic group can stretch symmetrically (in-phase) or asymmetrically (out-of-phase). Similar to hydrogen bonding, these vibrations also influence the shape of the IR peak. Generally, asymmetric stretching frequencies are higher than symmetric stretching frequencies. For example, primary amines exhibit two distinct IR peaks between 3300–3500 cm−1 corresponding to the symmetric and asymmetric N-H stretching, while secondary amines exhibit a single stretching vibration...
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...
¹H NMR Signal Integration: Overview00:58

¹H NMR Signal Integration: Overview

The intensity of a signal, which can be represented by the area under the peak, depends on the number of protons contributing to that signal. The area under each peak is shown as a vertical line called an integral, with the integral value listed under it, as seen in the proton NMR spectrum of benzyl acetate. Each integral value is divided by the smallest integral value to obtain the ratio of the number of protons producing each signal. The ratio reveals the relative number of protons and not...
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.
Mass Spectrum01:23

Mass Spectrum

A mass spectrum is the graphical representation of the relative abundance of the charged fragments in an analyte plotted against their mass-to-charge ratio (m/z). The plot's x-axis represents the ratio of the mass of the charged fragment to the number of charges it carries. The y axis of the plot represents the relative abundance of each charged species. The relative abundance is calculated from the signal intensity of each charged species recorded at the detector. The most intense signal (the...

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Related Experiment Video

Updated: Jul 11, 2026

Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR
14:44

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

Published on: December 16, 2013

Protein assignments without peak lists using higher-order spectra.

Gregory Benison1, Donald S Berkholz, Elisar Barbar

  • 1Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR 97331, USA. gbenison@gmail.com

Journal of Magnetic Resonance (San Diego, Calif. : 1997)
|October 9, 2007
PubMed
Summary

Visualizing higher-order spectra aids biomolecule assignment by identifying peak relationships directly from spectra. This method complements automated tools for protein structure determination.

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Analyzing Large Protein Complexes by Structural Mass Spectrometry
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Analyzing Large Protein Complexes by Structural Mass Spectrometry

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Related Experiment Videos

Last Updated: Jul 11, 2026

Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR
14:44

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

Published on: December 16, 2013

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

Published on: September 17, 2017

Analyzing Large Protein Complexes by Structural Mass Spectrometry
15:35

Analyzing Large Protein Complexes by Structural Mass Spectrometry

Published on: June 19, 2010

Area of Science:

  • Biochemistry
  • Structural Biology
  • Spectroscopy

Background:

  • Automated methods for biomolecule peak list generation and manipulation have advanced.
  • Direct spectral analysis offers advantages over algorithms for identifying peak relationships, especially with noise and overlap.
  • Existing tools for peak picking and automated assignment can be augmented.

Purpose of the Study:

  • To introduce higher-order spectra as a tool for direct visual identification of peak relationships in biomolecular spectra.
  • To demonstrate the utility of higher-order spectra for solving the biomolecular assignment problem.
  • To provide a software tool for generating and visualizing higher-order spectra.

Main Methods:

  • Development and application of higher-order spectra for analyzing spectral data.
  • Direct visual examination of contour plots to identify correlated peaks.
  • Utilizing the "burrow-owl" program for generating and displaying higher-order spectra.

Main Results:

  • Higher-order spectra enable direct visual identification of peak relationships, similar to searching peak lists.
  • The technique was successfully applied to achieve novel, complete sequential assignment of two proteins: AhpFn and IC74(84-143).
  • The "burrow-owl" software facilitates the generation and display of these higher-order spectra.

Conclusions:

  • Higher-order spectra offer a valuable complementary approach to automated methods for biomolecular assignment.
  • Direct visual inspection of higher-order spectra can effectively identify crucial peak correlations.
  • This technique enhances the toolkit available for tackling complex spectral assignment challenges in structural biology.