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

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...
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...
Interaction of EM Radiation with Matter: Spectroscopy01:12

Interaction of EM Radiation with Matter: Spectroscopy

Electromagnetic (EM) radiation can be considered an oscillating electric and magnetic field propagating through a medium that can interact with matter in its path. The electric field in the radiation can interact with electrical charges in the atoms or molecules in the matter. On the other hand, the magnetic field can interact with the magnetic field in the atomic nucleus. The study of the interaction between electromagnetic radiation and matter is termed spectroscopy. Spectroscopy is the study...
Insensitive Nuclei Enhanced by Polarization Transfer (INEPT)01:15

Insensitive Nuclei Enhanced by Polarization Transfer (INEPT)

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...
Double Resonance Techniques: Overview01:12

Double Resonance Techniques: Overview

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...
NMR Spectrometers: Resolution and Error Correction01:14

NMR Spectrometers: Resolution and Error Correction

When magnetic nuclei in a sample achieve resonance and undergo relaxation, the signal detected in NMR is an approximately exponential free induction decay. Fourier transform of an exponential decay yields a Lorentzian peak in the frequency domain. Lorentzian peaks in an NMR spectrum are defined by their amplitude, full width at half maximum, and position, where the peak width is governed by the spin-spin relaxation time alone. In real experiments, however, the applied magnetic field is rendered...

You might also read

Related Articles

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

Sort by
Same author

Severe lymphomatoid papulosis with lymphadenopathy, facial involvement, and a pathogenic STAT3 variant.

JAAD case reports·2026
Same author

Electrochemical characterization of photo-driven hole-scavenging by cadmium sulfide quantum dot-nitrogenase biohybrid complexes.

Bioelectrochemistry (Amsterdam, Netherlands)·2026
Same author

Sulfite Is Not Required for N<sub>2</sub> Reduction Catalyzed by Mo-Nitrogenase.

Journal of the American Chemical Society·2026
Same author

Mechanistic Insights into Dinitrogen Reduction to Ammonia in Light-Controlled Nanocrystal:Nitrogenase Complexes.

Accounts of chemical research·2026
Same author

Resurrected nitrogenases recapitulate canonical N-isotope biosignatures over two billion years.

Nature communications·2026
Same author

Carbon monoxide chemistry of α-V70I Mo-nitrogenase: Evidence from EPR- and IR-monitored photolysis - or, what a difference a methyl makes.

Journal of inorganic biochemistry·2026
Same journal

Tracking Synthetic Adhesins on Bacterial Surfaces with Immunofluorescence Microscopy.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Post-Selection Methods for Analyzing mRNA Display Selections and Optimization of Hits.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

High-Performance Computing in Tandem Mass Spectrometry (MS/MS) Peptide Identification.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Engineering and Adapting Disulfide-Containing Proteins to Enable Intracellular Functionality.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

AI-Driven Protein Research: From Prediction to Design.

Methods in molecular biology (Clifton, N.J.)·2026
Same journal

Methods for the In Vitro Selection of Protein and Peptide Libraries Using mRNA Display.

Methods in molecular biology (Clifton, N.J.)·2026
See all related articles

Related Experiment Video

Updated: May 30, 2026

Rapid Scan Electron Paramagnetic Resonance Opens New Avenues for Imaging Physiologically Important Parameters In Vivo
08:01

Rapid Scan Electron Paramagnetic Resonance Opens New Avenues for Imaging Physiologically Important Parameters In Vivo

Published on: September 26, 2016

Electron paramagnetic resonance spectroscopy.

Karamatullah Danyal1, Zhi-Yong Yang, Lance C Seefeldt

  • 1Department of Chemistry and Biochemistry, Utah State University, Logan, UT 84322, USA. k.danyal@aggiemail.usu.edu

Methods in Molecular Biology (Clifton, N.J.)
|August 12, 2011
PubMed
Summary
This summary is machine-generated.

Electron paramagnetic resonance (EPR) spectroscopy is crucial for studying nitrogenase, a key enzyme. This method analyzes the enzyme's metal clusters and substrate interactions, advancing nitrogenase research.

More Related Videos

Use of Electron Paramagnetic Resonance in Biological Samples at Ambient Temperature and 77 K
06:45

Use of Electron Paramagnetic Resonance in Biological Samples at Ambient Temperature and 77 K

Published on: January 11, 2019

Exploring the Radical Nature of a Carbon Surface by Electron Paramagnetic Resonance and a Calibrated Gas Flow
10:34

Exploring the Radical Nature of a Carbon Surface by Electron Paramagnetic Resonance and a Calibrated Gas Flow

Published on: April 24, 2014

Related Experiment Videos

Last Updated: May 30, 2026

Rapid Scan Electron Paramagnetic Resonance Opens New Avenues for Imaging Physiologically Important Parameters In Vivo
08:01

Rapid Scan Electron Paramagnetic Resonance Opens New Avenues for Imaging Physiologically Important Parameters In Vivo

Published on: September 26, 2016

Use of Electron Paramagnetic Resonance in Biological Samples at Ambient Temperature and 77 K
06:45

Use of Electron Paramagnetic Resonance in Biological Samples at Ambient Temperature and 77 K

Published on: January 11, 2019

Exploring the Radical Nature of a Carbon Surface by Electron Paramagnetic Resonance and a Calibrated Gas Flow
10:34

Exploring the Radical Nature of a Carbon Surface by Electron Paramagnetic Resonance and a Calibrated Gas Flow

Published on: April 24, 2014

Area of Science:

  • Biochemistry
  • Bioinorganic Chemistry
  • Spectroscopy

Background:

  • Nitrogenase research relies heavily on Electron Paramagnetic Resonance (EPR) spectroscopy.
  • The enzyme contains three essential metalloclusters: the Fe protein [4Fe-4S] cluster, the MoFe protein P-cluster, and the FeMo-cofactor.
  • These metalloclusters exhibit EPR spectra in specific paramagnetic states, making them amenable to spectroscopic analysis.

Purpose of the Study:

  • To detail the methods for analyzing the three metal clusters of nitrogenase using EPR spectroscopy.
  • To describe techniques for trapping and characterizing substrate-derived intermediates within the nitrogenase active site.
  • To highlight the utility of EPR and other magnetic resonance techniques in understanding nitrogenase function.

Main Methods:

  • Utilizing Electron Paramagnetic Resonance (EPR) spectroscopy to analyze nitrogenase metalloclusters.
  • Employing EPR to monitor changes in electronic properties and redox states of metal clusters.
  • Implementing methods for trapping substrate-derived intermediates for magnetic resonance characterization.

Main Results:

  • EPR spectroscopy provides detailed insights into the electronic properties of nitrogenase's metal clusters.
  • The technique allows for definitive observation of redox state changes within the clusters.
  • Substrate binding events can be effectively probed by monitoring alterations in EPR spectra.

Conclusions:

  • EPR spectroscopy is an indispensable tool for the comprehensive study of nitrogenase.
  • The described methods enable detailed characterization of nitrogenase's metal centers and catalytic intermediates.
  • This approach significantly advances our understanding of nitrogenase's mechanism and function.