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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)

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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...
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Proton (¹H) NMR: Chemical Shift01:07

Proton (¹H) NMR: Chemical Shift

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Organic molecules primarily contain carbon and hydrogen atoms. While all the hydrogen isotopes are NMR-active, protium or hydrogen-1 is the most abundant. It has a significant energy separation between its nuclear spin states due to its large gyromagnetic ratio. As per Boltzmann's distribution, an increase in the energy separation implies a greater excess population of nuclei available for excitation, resulting in a strong NMR absorption signal.
Absorption signals of all the protium nuclei...
1.7K
Chemical Shift: Internal References and Solvent Effects01:17

Chemical Shift: Internal References and Solvent Effects

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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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Applications Of NMR In Biology01:25

Applications Of NMR In Biology

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Nuclear magnetic resonance (NMR) spectroscopy is a very valuable analytical technique for researchers. It has been used for more than 50 years as an analytical tool. F. Bloch and E. Purcell formulated NMR in 1946 and won the 1952 Nobel Prize in Physics  for their work. Biological macromolecules such as proteins, nucleic acids, lipids, and organic molecules including pharmaceutical compounds, can be studied using this versatile tool that exploits the magnetic properties of certain nuclei.
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¹H NMR of Labile Protons: Temporal Resolution01:10

¹H NMR of Labile Protons: Temporal Resolution

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Protons bonded to heteroatoms such as nitrogen and oxygen exhibit a range of chemical shift values. This is due to the varying degree of hydrogen bonding between the proton and the heteroatom in other molecules. The extent of hydrogen bonding affects the electron density around the proton, thereby giving different chemical shift values for the protons in the proton NMR spectrum.
The –OH proton in alcohols typically appears in the range of δ 2 to 5 ppm but can vary depending on the specific...
1.2K
¹H NMR of Labile Protons: Deuterium (²H) Substitution00:48

¹H NMR of Labile Protons: Deuterium (²H) Substitution

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This lesson illustrates the role of deuterium substitution in simplifying the NMR spectrum of compounds comprising labile protons. One method employed is the use of deuterium. Amongst the three isotopes of hydrogen, deuterium (2H) has a nucleus composed of one proton and one neutron. When the D2O solvent is added to a pure dry ethanol solution, its labile proton is substituted with deuterium.
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Related Experiment Video

Updated: Jul 12, 2025

Methods to Identify the NMR Resonances of the 13C-Dimethyl N-terminal Amine on Reductively Methylated Proteins
13:59

Methods to Identify the NMR Resonances of the 13C-Dimethyl N-terminal Amine on Reductively Methylated Proteins

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Protein methylation characterization using NMR without isotopic labeling.

Zhongpei Fang1, Tao Huang2, Xin Chai1

  • 1Key Laboratory of Magnetic Resonance in Biological Systems, State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, National Center for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Innovation Academy for Precision Measurement of Science and Technology, Chinese Academy of Sciences, Wuhan, 430071, China; University of Chinese Academy of Sciences, Beijing, 100049, China.

Talanta
|October 20, 2023
PubMed
Summary
This summary is machine-generated.

Protein methylation, crucial in epigenetics, can now be easily characterized without isotopic labeling. This new method reveals varying histone H3 methylation in cancer, aiding therapeutic development.

Keywords:
Costly savingIsotope freeNMR spectroscopyProtein methylation

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A Mass Spectrometry-Based Proteomics Approach for Global and High-Confidence Protein R-Methylation Analysis
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Exploring the Arginine Methylome by Nuclear Magnetic Resonance Spectroscopy
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Area of Science:

  • Biochemistry
  • Epigenetics
  • Oncology

Background:

  • Protein methylation is vital in epigenetics and a target for cancer therapies.
  • Current methods for measuring methylation, like NMR, can be costly and limited.
  • Characterizing methylation is key to understanding methyltransferase function and developing inhibitors.

Purpose of the Study:

  • To develop a simple, non-isotopic labeling method for characterizing protein methylation and demethylation.
  • To assess histone H3 methylation patterns in various mouse cell and tissue lysates, including cancerous ones.

Main Methods:

  • Utilized a four-quantum filter 1H-13C experiment to selectively detect methyl groups.
  • Applied the method to analyze histone H3 methylation in lysates from healthy, cancerous, and tumor tissues.
  • No isotopic labeling of protein or methyl donors was required.

Main Results:

  • Successfully observed mono- and dimethylation of histone H3 in all tested mouse lysates.
  • Found significantly lower H3 methylation rates and levels in cervical and breast tumor lysates compared to cancerous and healthy cells.
  • Demonstrated variability in H3 methylation patterns across different cell types, tissues, and cancer stages.

Conclusions:

  • The developed method offers a straightforward way to characterize protein methylation and demethylation.
  • Histone H3 methylation patterns vary significantly in cancer, offering insights into disease mechanisms.
  • This technique holds potential for developing targeted cancer interventions by analyzing methylation features.