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Related Concept Videos

2D NMR: Overview of Heteronuclear Correlation Techniques01:18

2D NMR: Overview of Heteronuclear Correlation Techniques

683
Heteronuclear correlation spectroscopy is an analytical technique that investigates the coupling between different types of nuclei, often a proton and an X-nucleus, such as carbon-13 or nitrogen-15. This method is commonly used in nuclear magnetic resonance (NMR) spectroscopy to gain insights into complex chemical compounds' structural and compositional aspects. A typical heteronuclear correlation spectrum displays X-nucleus chemical shifts on one axis and a proton spectrum on the other...
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2D NMR: Overview of Homonuclear Correlation Techniques01:16

2D NMR: Overview of Homonuclear Correlation Techniques

569
Homonuclear correlation spectroscopy (COSY) is a powerful technique used in Nuclear Magnetic Resonance (NMR) spectroscopy to study the correlations between nuclei of the same type within a molecule. It provides information about scalar couplings between adjacent nuclei, which helps determine connectivity and structural information. There are several COSY variants, each with its unique strengths and experimental parameters.
COSY90 is the standard two-dimensional (2D) COSY experiment that...
569
2D NMR: Heteronuclear Single-Quantum Correlation Spectroscopy (HSQC)01:19

2D NMR: Heteronuclear Single-Quantum Correlation Spectroscopy (HSQC)

1.3K
Heteronuclear single-quantum correlation spectroscopy (HSQC) is a 2D NMR technique that reveals one-bond correlations between hydrogen and a heteronucleus. The HSQC experiment is similar to the heteronuclear correlation experiment (HETCOR) but is more sensitive. In the HSQC spectrum, the proton chemical shift is plotted on the horizontal F2 axis, while the 13C chemical shift is plotted on the vertical F1 axis. The corresponding proton and 13C spectra are also shown. The HSQC contour plot does...
1.3K
2D NMR: Homonuclear Correlation Spectroscopy (COSY)01:06

2D NMR: Homonuclear Correlation Spectroscopy (COSY)

1.8K
Homonuclear correlation spectroscopy, or COSY, is a 2-dimensional NMR technique that provides information about coupled protons. Typically, the geminal and vicinal coupling are observed. For example, consider the COSY spectrum of ethyl acetate, where its 1D proton NMR spectrum is plotted along the vertical and horizontal axes with their corresponding chemical shift scale. Three spots on the diagonal corresponding to the three peaks in the 1D proton spectrum are called diagonal peaks. The COSY...
1.8K
¹H NMR: Interpreting Distorted and Overlapping Signals01:02

¹H NMR: Interpreting Distorted and Overlapping Signals

1.4K
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...
1.4K
Two-Dimensional (2D) NMR: Overview01:12

Two-Dimensional (2D) NMR: Overview

1.4K
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....
1.4K

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Two-dimensional NMR lineshape analysis of single, multiple, zero and double quantum correlation experiments.

Christopher A Waudby1, Margaux Ouvry2, Ben Davis3

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Summary

This study classifies chemical exchange regimes in 2D NMR, detailing differences between HSQC and HMQC experiments. New experiments like SOFAST-H(Z/D)QC and BEST-ZQ-TROSY improve characterization of molecular interactions.

Keywords:
Chemical exchangeDouble-quantumLineshape analysisMultiple-quantumTitrationsTwo-dimensional NMRZero-quantum

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Area of Science:

  • Biophysical Chemistry
  • Structural Biology
  • Nuclear Magnetic Resonance (NMR) Spectroscopy

Background:

  • Nuclear Magnetic Resonance (NMR) spectroscopy is crucial for studying chemical exchange and molecular interactions through titration experiments.
  • Understanding chemical exchange is vital, but 2D NMR experiments present complexities beyond 1D methods.
  • Existing 2D NMR techniques can suffer from limitations like exchange broadening, impacting data quality.

Purpose of the Study:

  • To systematically classify chemical exchange regimes in two-dimensional (2D) NMR spectra.
  • To compare exchange broadening effects in Heteronuclear Single Quantum Coherence (HSQC) and Heteronuclear Multiple Quantum Coherence (HMQC) experiments.
  • To introduce and validate novel NMR experiments for enhanced characterization of molecular interactions.

Main Methods:

  • Development of a systematic classification for 2D NMR chemical exchange regimes.
  • Comparative analysis of HSQC, HMQC, Heteronuclear Zero Quantum Coherence (HZQC), and Heteronuclear Double Quantum Coherence (HDQC) experiments.
  • Introduction of the SOFAST-H(Z/D)QC and BEST-ZQ-TROSY experiments for improved sensitivity and data acquisition.

Main Results:

  • Identified distinct chemical exchange regimes in 2D NMR, highlighting greater exchange broadening in HMQC spectra.
  • Demonstrated that HZQC and HDQC experiments serve as viable alternatives to HMQC, mitigating broadening issues.
  • Successfully applied the new experiments to characterize the interaction between the Hsp90 N-terminal domain and a small molecule ligand.

Conclusions:

  • The classification and new experiments enhance confidence in fitted dissociation constants and rates by enabling independent analysis of various 2D NMR data.
  • Joint analysis of data from HSQC, HMQC, HZQC, and HDQC experiments can improve sensitivity for complex multi-state interaction mechanisms.
  • These advancements offer powerful tools for detailed structural and dynamic characterization of biomolecular interactions.