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Related Concept Videos

¹³C NMR: ¹H–¹³C Decoupling01:04

¹³C NMR: ¹H–¹³C Decoupling

The probability of having two carbon-13 atoms next to each other is negligible because of the low natural abundance of carbon-13. Consequently, peak splitting due to carbon-carbon spin-spin coupling is not observed in spectra. However, protons up to three sigma bonds away split the carbon signal according to the n+1 rule, resulting in complicated spectra.
A broadband decoupling technique is used to simplify these complex, sometimes overlapping, signals. Broadband decoupling relies on a...
Mass Spectrometry: Isotope Effect01:13

Mass Spectrometry: Isotope Effect

Most elements exist in nature as a mixture of isotopes. The isotopes differ in weight due to their respective number of neutrons. The molecular weight of a molecule is different depending on the specific isotope of its elements involved. As a result, the mass spectrum of the molecule exhibits peaks from the same fragment at multiple positions. The positions of these mass signals depend on the mass differences between isotopes. Furthermore, the intensity of these signals is dependent on the...
Conservation of Protein Domains02:26

Conservation of Protein Domains

Protein domains are small structurally independent units that are part of a single amino acid chain.  Although these domains are often structurally independent, they may rely on synergistic effects to perform their functions as part of a larger protein. Protein domains may be conserved within the same organism, as well as across different organisms.
A limited set of protein domains often duplicate and recombine during evolution. These domains can be organized in different combinations to form...
Conservation of Protein Domains Over Different Proteins02:26

Conservation of Protein Domains Over Different Proteins

Protein domains are small structurally independent units that are part of a single amino acid chain.  Although these domains are often structurally independent, they may rely on synergistic effects to perform their functions as part of a larger protein. Protein domains may be conserved within the same organism, as well as across different organisms.
A limited set of protein domains often duplicate and recombine during evolution. These domains can be organized in different combinations to form...
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...
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...

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Related Experiment Video

Updated: May 31, 2026

Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling
11:53

Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling

Published on: July 1, 2014

Isotope Decluttering Reduces Spectral Complexity while Maintaining Protein Structure.

Sneha S Hakke1, Willem E M Noteborn2, Birol Cabukusta1

  • 1Maastricht Multi-Modal Molecular Imaging Institute (M4i), Maastricht University, 6229 ER Maastricht, The Netherlands.

Analytical Chemistry
|May 29, 2026
PubMed
Summary
This summary is machine-generated.

Isotope depletion simplifies mass spectrometry (MS) analysis of large proteins and complexes by reducing isotopic complexity. This method enhances signal-to-noise ratio for accurate mass measurement and maintains structural integrity for detailed analysis.

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Last Updated: May 31, 2026

Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling
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Published on: July 1, 2014

NMR 15N Relaxation Experiments for the Investigation of Picosecond to Nanoseconds Structural Dynamics of Proteins
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Neutron Crystallography Data Collection and Processing for Modelling Hydrogen Atoms in Protein Structures
10:10

Neutron Crystallography Data Collection and Processing for Modelling Hydrogen Atoms in Protein Structures

Published on: December 1, 2020

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Structural Biology

Background:

  • Accurate mass determination is crucial in mass spectrometry (MS) for molecular identification.
  • Heavier isotopes increase spectral complexity and reduce signal-to-noise ratio (SNR), especially for large proteins and complexes.
  • Current methods struggle with spectral complexity in large biomolecules.

Purpose of the Study:

  • To investigate the impact of isotope depletion on mass spectrometry analysis of large protein complexes.
  • To assess the structural integrity of isotope-depleted proteins and complexes.
  • To evaluate isotope depletion as a method for improving MS-based protein identification.

Main Methods:

  • Expression and purification of *Mycobacterium tuberculosis* protein complexes (EsxAB and BfrB) under isotope-depleted conditions.
  • Application of isotope depletion to bacterioferritin B (BfrB), the largest reported complex analyzed under native conditions.
  • Analysis of structural integrity using single-particle cryo-electron microscopy (cryo-EM).

Main Results:

  • Isotope-depleted proteins and complexes exhibited simplified mass spectra with reduced isotopic distribution.
  • Significant increase in the SNR of the monoisotopic peak was observed.
  • Improved protein sequence coverage was achieved using native top-down MS.
  • Cryo-EM confirmed that isotope depletion preserves protein structural integrity at atomic resolution.

Conclusions:

  • Isotope depletion effectively simplifies mass spectra and enhances SNR for large proteins and complexes.
  • This technique improves accuracy in mass measurement and protein identification via MS.
  • Isotope depletion is a viable method for high-accuracy MS analysis without compromising protein structure.