Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Electron Paramagnetic Resonance (EPR) Spectroscopy: Organic Radicals01:17

Electron Paramagnetic Resonance (EPR) Spectroscopy: Organic Radicals

3.3K
Ideally, an unpaired electron shows a single peak in the EPR spectrum due to the transition between the two spin energy states. However, coupling interactions can occur between the spins of the unpaired electron and any neighboring spin-active nuclei. This hyperfine coupling results in hyperfine splitting, where the EPR signal is split into multiplets. The signals split into 2nI + 1 peaks, where n is the number of equivalent nuclei and I is the nuclear spin. These splitting patterns provide...
3.3K
Double Resonance Techniques: Overview01:12

Double Resonance Techniques: Overview

705
Double resonance techniques in Nuclear Magnetic Resonance (NMR) spectroscopy involve the simultaneous application of two different frequencies or radiofrequency pulses to manipulate and observe two distinct nuclear spins. One important application of double resonance is spin decoupling, which selectively suppresses coupling with one type of nucleus while observing the NMR signal from another nucleus, simplifying the spectrum and enhancing resolution.
Spin decoupling is usually achieved by...
705
¹³C NMR: Distortionless Enhancement by Polarization Transfer (DEPT)01:20

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

1.6K
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...
1.6K
Insensitive Nuclei Enhanced by Polarization Transfer (INEPT)01:15

Insensitive Nuclei Enhanced by Polarization Transfer (INEPT)

1.0K
Insensitive Nuclei Enhanced by Polarization Transfer (INEPT) is an advanced Nuclear Magnetic Resonance (NMR) technique specifically designed to detect and enhance the signals of low-abundance nuclei, such as carbon-13 and nitrogen-15, in small molecules. The fundamental principle behind INEPT is the transfer of polarization from a more abundant and highly polarizable nucleus, typically hydrogen-1, to the low-abundance nucleus of interest. This process effectively boosts the NMR signal of the...
1.0K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Immobilization-mediated algae-bacteria interactions improve swine wastewater treatment and microalgal biomass energy conversion: Performance, correlation and microscopic mechanisms.

Journal of environmental management·2026
Same author

Unraveling Synthetase's Mode of Action: The Pyrrolysyl-tRNA Synthetase Dimer Uses Secondary Binding Sites in the Cell.

Angewandte Chemie (International ed. in English)·2026
Same author

Study on the association between dietary inflammatory index and sarcopenia in older adults.

BMC geriatrics·2026
Same author

Effect of postoperative coffee consumption on gastrointestinal function after cesarean section, a systematic review and meta-analysis of randomized controlled trials.

BMC public health·2026
Same author

Effects of single-frequency ultrasonic modification on starch structure and digestive properties: A comprehensive meta-analysis and acoustic simulation.

Food chemistry: X·2026
Same author

Eu<sup>2+</sup> and Ce<sup>3+</sup>/Eu<sup>2+</sup> co-doped Ba<sub>2</sub>La<sub>3</sub>(SiO<sub>4</sub>)<sub>2</sub>(PO<sub>4</sub>)O Phosphors: Luminescence Properties and Applications in LEDs and Temperature Sensing.

Journal of fluorescence·2025
Same journal

Neutral Amphiphiles Boost Transfection Efficiency and Reduce Inflammation in Polymer Micelle-Mediated mRNA Delivery.

Bioconjugate chemistry·2026
Same journal

Surfactant-Mediated Buchwald-Hartwig Coupling of Aliphatic Amines for the Synthesis of DNA-Encoded Libraries.

Bioconjugate chemistry·2026
Same journal

Artificial Intelligence for Discovery in Life Sciences.

Bioconjugate chemistry·2026
Same journal

Iron Single Atom Nanozyme-Mediated GPX4 Inhibitor Delivery for Self-Enhanced Ferroptosis.

Bioconjugate chemistry·2026
Same journal

SpyCatcher-Engineered Ferritin Nanocages Enable Dual-Receptor Targeting for Enhanced Glioma Therapy.

Bioconjugate chemistry·2026
Same journal

One-Pot Synthesis of Functionalized Coumarin Fluorophores Enables Rapid Access to Live-Cell Bioorthogonal Labeling and Microenvironmental Sensing Agents.

Bioconjugate chemistry·2026
See all related articles

Related Experiment Video

Updated: Jan 16, 2026

Site Directed Spin Labeling and EPR Spectroscopic Studies of Pentameric Ligand-Gated Ion Channels
11:19

Site Directed Spin Labeling and EPR Spectroscopic Studies of Pentameric Ligand-Gated Ion Channels

Published on: July 4, 2016

11.0K

Cysteine-Targeting Gd-Based Spin Label and Its Application in Electron Paramagnetic Resonance Spectroscopy.

Xuemei Yao1, Eliane Landwehr2, Mian Qi1

  • 1Faculty of Chemistry and Center of Molecular Materials (CM2), Bielefeld University, Universitätsstraße 25, Bielefeld 33615, Germany.

Bioconjugate Chemistry
|September 26, 2025
PubMed
Summary
This summary is machine-generated.

Researchers developed a faster method for attaching paramagnetic labels to proteins using a pyrimidine-based reaction. This technique enhances protein structure determination via electron paramagnetic resonance (EPR) and nuclear magnetic resonance (NMR) spectroscopy.

More Related Videos

Paramagnetic Relaxation Enhancement for Detecting and Characterizing Self-Associations of Intrinsically Disordered Proteins
07:24

Paramagnetic Relaxation Enhancement for Detecting and Characterizing Self-Associations of Intrinsically Disordered Proteins

Published on: September 23, 2021

2.2K
Purification and Reconstitution of TRPV1 for Spectroscopic Analysis
11:53

Purification and Reconstitution of TRPV1 for Spectroscopic Analysis

Published on: July 3, 2018

8.4K

Related Experiment Videos

Last Updated: Jan 16, 2026

Site Directed Spin Labeling and EPR Spectroscopic Studies of Pentameric Ligand-Gated Ion Channels
11:19

Site Directed Spin Labeling and EPR Spectroscopic Studies of Pentameric Ligand-Gated Ion Channels

Published on: July 4, 2016

11.0K
Paramagnetic Relaxation Enhancement for Detecting and Characterizing Self-Associations of Intrinsically Disordered Proteins
07:24

Paramagnetic Relaxation Enhancement for Detecting and Characterizing Self-Associations of Intrinsically Disordered Proteins

Published on: September 23, 2021

2.2K
Purification and Reconstitution of TRPV1 for Spectroscopic Analysis
11:53

Purification and Reconstitution of TRPV1 for Spectroscopic Analysis

Published on: July 3, 2018

8.4K

Area of Science:

  • Biochemistry
  • Chemical Biology
  • Spectroscopy

Background:

  • Selective labeling of cysteine residues in peptides and proteins is crucial for structural studies.
  • Existing methods using Michael addition with 4-vinylpyridine are often too slow for broad application.
  • Paramagnetic labels are essential for techniques like electron paramagnetic resonance (EPR) and nuclear magnetic resonance (NMR) spectroscopy.

Purpose of the Study:

  • To develop a faster and highly selective method for anchoring paramagnetic labels to cysteine residues.
  • To evaluate the utility of a novel pyrimidine-based labeling agent for protein structure determination.
  • To assess the impact of replacing pyridine with pyrimidine on reaction kinetics and EPR properties.

Main Methods:

  • Michael addition reaction between cysteine-containing biomolecules and 4-vinylpyrimidine-based metal complexes (4-vinyl-PymiMTA-Ln).
  • Application of the labeling agent to cysteine, cysteine-containing oligoproline, and cysteine-containing thioredoxin.
  • Characterization of the resulting spin labels using EPR and double electron-electron resonance (DEER) spectroscopy.

Main Results:

  • The reaction of 4-vinylpyrimidine derivatives with thiols is significantly faster than their pyridine counterparts.
  • The chemoselectivity of the reaction is maintained, forming reduction-resistant linkages.
  • The PymiMTA-Gd complex proved effective as a spin label for distance determination using DEER spectroscopy.
  • EPR spectra and relaxation times of pyrimidine- and pyridine-based labels were found to be similar.

Conclusions:

  • Replacing pyridine with pyrimidine in metal complexes creates fast-reacting, chemoselective spin labels for biomolecules.
  • These novel labels retain favorable EPR spectroscopical properties, enabling advanced structural analysis.
  • This strategy offers a versatile approach for developing new labeling agents for EPR and NMR studies.