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Related Concept Videos

2D NMR: Overview of Heteronuclear Correlation Techniques01:18

2D NMR: Overview of Heteronuclear Correlation Techniques

179
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...
179
2D NMR: Heteronuclear Single-Quantum Correlation Spectroscopy (HSQC)01:19

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

716
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...
716
2D NMR: Overview of Homonuclear Correlation Techniques01:16

2D NMR: Overview of Homonuclear Correlation Techniques

198
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...
198
¹H NMR: Interpreting Distorted and Overlapping Signals01:02

¹H NMR: Interpreting Distorted and Overlapping Signals

1.0K
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...
1.0K
2D NMR: Homonuclear Correlation Spectroscopy (COSY)01:06

2D NMR: Homonuclear Correlation Spectroscopy (COSY)

1.1K
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...
1.1K

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Mass Spectrometry and Luminogenic-based Approaches to Characterize Phase I Metabolic Competency of In Vitro Cell Cultures
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Heme Spin Distribution in the Substrate-Free and Inhibited Novel CYP116B5hd: A Multifrequency Hyperfine Sublevel

Antonino Famulari1,2, Danilo Correddu3, Giovanna Di Nardo3

  • 1Departamento de Física de la Materia Condensada, Universidad de Zaragoza, C/Pedro Cerbuna 12, 50009 Zaragoza, Spain.

Molecules (Basel, Switzerland)
|January 26, 2024
PubMed
Summary

This study details the electronic structure of CYP116B5hd, a cytochrome P450 enzyme, using hyperfine spectroscopy. It reveals active site details and imidazole binding effects, crucial for understanding enzyme function and drug development.

Keywords:
CYP450EPR spectroscopyHYSCOREhyperfine interactionsimidazole bindinglow-spin hemeproteinmultifrequency EPRperoxygenasequadrupole interaction

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Area of Science:

  • Biochemistry and Molecular Biology
  • Enzymology
  • Spectroscopy

Background:

  • Cytochrome P450 (CYP) enzymes are vital monooxygenases with diverse catalytic capabilities.
  • CYP116B5hd exhibits notable peracid resistance, making it valuable for fine chemical synthesis.
  • Understanding CYP active site structure and inhibition mechanisms is critical for drug discovery, particularly in anti-cancer therapies.

Purpose of the Study:

  • To characterize the electronic structure of the heme iron in CYP116B5hd using multifrequency hyperfine spectroscopy.
  • To elucidate the active site geometry and identify key interacting protons.
  • To investigate the effects of imidazole binding on CYP116B5hd, modeling P450 inhibition.

Main Methods:

  • Multifrequency (X- and Q-band) hyperfine spectroscopy was employed to study CYP116B5hd.
  • HYSCORE experiments were utilized to detect and analyze hyperfine interactions.
  • Analysis of gyromagnetic, hyperfine, and quadrupole tensors provided insights into active site structure and electronic properties.

Main Results:

  • Detailed characterization of the heme iron's electronic structure in the resting state of CYP116B5hd.
  • Localization of water and cysteine ligand protons relative to the heme iron's magnetic axes.
  • Imidazole binding was shown to induce axial coordination and a low-spin Fe(III) state, with two distinct binding geometries identified.

Conclusions:

  • The study provides precise structural details of the CYP116B5hd active site, including orbital orientation.
  • Imidazole binding does not significantly alter the electronic structure of the iron but affects coordination geometry.
  • Findings contribute to understanding P450 enzyme mechanisms and developing targeted inhibitors for therapeutic applications.