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

¹H NMR: Interpreting Distorted and Overlapping Signals01:02

¹H NMR: Interpreting Distorted and Overlapping Signals

926
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...
926
¹³C NMR: Distortionless Enhancement by Polarization Transfer (DEPT)01:20

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

952
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...
952
Two-Dimensional (2D) NMR: Overview01:12

Two-Dimensional (2D) NMR: Overview

564
The 1D NMR spectrum of large and complex molecules like natural products has complicated splitting patterns and overlapping signals, which can be easily interpreted using 2-dimensional (2D) NMR. Unlike 1D NMR, 2D NMR has two frequency axes that provide the coupling information between the nucleus A and nucleus B in a molecule. The process from which 2D spectra are obtained has four steps.
The first step is the preparation period, during which nucleus A is excited with a radiofrequency pulse....
564
¹H NMR of Conformationally Flexible Molecules: Temporal Resolution00:52

¹H NMR of Conformationally Flexible Molecules: Temporal Resolution

781
At room temperature, the chair conformer of cyclohexane undergoes rapid ring flipping between two equivalent chair conformers at a rate of approximately 105 times per second. These two chair conformers are in equilibrium. The rapid ring flipping results in the interconversion of the axial proton to an equatorial proton and an equatorial to the axial proton. Such interconversions are too rapid and cannot be detected on the NMR timescale. Hence, the NMR spectrometer cannot distinguish between the...
781
Intrinsically Disordered Proteins02:18

Intrinsically Disordered Proteins

17.6K
Intrinsically disordered proteins are a group of proteins that do not fold into specific three-dimensional structures. Their structural flexibility allows them to complement ordered proteins to perform functions that are inaccessible to rigid structures. They are more common in eukaryotes than prokaryotes and may either be exclusively intrinsically disordered or hybrid proteins, consisting of a mix of ordered and disordered regions. The absence of a rigid structure in these proteins can be...
17.6K
NMR Spectroscopy Of Amines01:19

NMR Spectroscopy Of Amines

8.3K
In proton NMR spectroscopy, primary amines and secondary amines showcase their N–H protons as a broad signal in the chemical shift range between δ 0.5 and 5 ppm. The exact position in this range depends on several factors, including sample concentration, hydrogen bonding, and the type of solvent used. Since amine protons undergo fast proton exchange in solution, the protons are labile and therefore do not participate in any splitting with adjacent protons. Thus, the observed peak is...
8.3K

You might also read

Related Articles

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

Sort by
Same author

A Rigid Supramolecular Solution to a Flexible Problem: A Multifunctional Calix[4]arene-Based Strategy to Prevent α‑Synuclein Toxicity.

ACS central science·2026
Same author

Molecular Interplay of Small Molecules and Calcium Ions with α-Synuclein Revealed by NMR and Molecular Dynamics Simulations.

ACS chemical neuroscience·2026
Same author

At the core of the interaction: Probing charged side chains in flexible protein regions with simultaneous nuclear magnetic resonance experiments.

Protein science : a publication of the Protein Society·2026
Same author

Oligomerization enables the selective targeting of an intrinsically disordered region by a small molecule.

Science advances·2026
Same author

Exploring Disordered Regions of Human Spliceosome Proteins.

The journal of physical chemistry letters·2026
Same author

<sup>15</sup>N optimal control pulses: an efficient approach to enhance heteronuclear-detected NMR experiments at high magnetic fields.

Journal of magnetic resonance (San Diego, Calif. : 1997)·2025

Related Experiment Video

Updated: May 13, 2025

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

1.7K

Decoding Order and Disorder in Proteins by NMR Spectroscopy.

Lorenzo Bracaglia1, Silvia Oliveti1, Isabella C Felli1

  • 1Department of Chemistry "Ugo Schiff" and Magnetic Resonance Center, University of Florence, Sesto Fiorentino 50019, Italy.

Journal of the American Chemical Society
|April 14, 2025
PubMed
Summary

Studying complex proteins like CREB binding protein (CBP) requires examining larger constructs. New NMR methods reveal interactions between intrinsically disordered regions (IDRs) and globular domains in CBP, offering deeper insights.

More Related Videos

Atomic Scale Structural Studies of Macromolecular Assemblies by Solid-state Nuclear Magnetic Resonance Spectroscopy
14:55

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

Published on: September 17, 2017

15.3K
Author Spotlight: Exploring Intrinsically Disordered Protein Dynamics Through NMR Relaxation Experiments
09:25

Author Spotlight: Exploring Intrinsically Disordered Protein Dynamics Through NMR Relaxation Experiments

Published on: November 1, 2024

1.6K

Related Experiment Videos

Last Updated: May 13, 2025

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

1.7K
Atomic Scale Structural Studies of Macromolecular Assemblies by Solid-state Nuclear Magnetic Resonance Spectroscopy
14:55

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

Published on: September 17, 2017

15.3K
Author Spotlight: Exploring Intrinsically Disordered Protein Dynamics Through NMR Relaxation Experiments
09:25

Author Spotlight: Exploring Intrinsically Disordered Protein Dynamics Through NMR Relaxation Experiments

Published on: November 1, 2024

1.6K

Area of Science:

  • Structural biology
  • Biochemistry
  • Molecular dynamics

Background:

  • Multidomain proteins, featuring globular domains and intrinsically disordered regions (IDRs), present challenges for traditional structural biology.
  • Isolating individual domains simplifies characterization but misses inter-domain interactions crucial for function.
  • Studying larger constructs is essential to understand the full complexity of multidomain proteins.

Purpose of the Study:

  • To characterize the TAZ4 construct of the CREB binding protein (CBP), which includes a globular domain and an IDR.
  • To develop and apply NMR techniques that leverage dynamic differences between domains for structural analysis.
  • To investigate potential crosstalk between the intrinsically disordered region and the globular domain within the TAZ4 construct.

Main Methods:

  • Utilized Nuclear Magnetic Resonance (NMR) spectroscopy, including 2D and 3D experiments.
  • Exploited differences in nuclear relaxation properties to select signals from distinct domains.
  • Performed sequence-specific assignment of the TAZ4 construct.

Main Results:

  • Successfully extended the sequence-specific assignment of the TAZ4 construct.
  • Demonstrated the feasibility of using NMR experiments sensitive to dynamics to study multidomain proteins.
  • Revealed evidence of crosstalk between the intrinsically disordered ID4 region and the globular TAZ2 domain.

Conclusions:

  • The study successfully characterized a challenging multidomain protein construct using advanced NMR techniques.
  • The findings highlight the importance of studying larger protein segments to understand inter-domain communication.
  • The observed crosstalk suggests functional interplay between disordered and ordered regions in CBP regulation.