Related Experiment Video

Updated: Jun 5, 2026

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

1H, 13C, and 15N backbone and side-chain chemical shift assignment of the staphylococcal MazF mRNA interferase

Valentina Zorzini1, Sarah Haesaerts, Ambrose Cheung

  • 1Structural Biology Brussels, Vrije Universiteit Brussel, Pleinlaan 2, 1050, Brussels, Belgium.

Biomolecular NMR Assignments
|January 8, 2011
PubMed

Insights

This study provides the first chemical shift assignments for the Staphylococcus aureus MazF toxin, a key component of bacterial stress response. These findings are crucial for understanding MazF function and developing novel antibacterial strategies.

Area of Science:

  • Biochemistry
  • Structural Biology
  • Microbiology

Background:

  • MazF proteins are sequence-specific ribonucleases involved in bacterial stress response, cleaving mRNA.
  • MazF toxins are regulated by antitoxins (MazE), and prolonged activation leads to cell death.
  • While MazEF systems in Gram-negative bacteria are well-studied, less is known about their Gram-positive counterparts.

Purpose of the Study:

  • To report the backbone and side-chain chemical shift assignments for the wild-type MazF toxin from Staphylococcus aureus.
  • To provide foundational data for future structural and functional studies of Gram-positive MazF toxins.

Main Methods:

  • Nuclear Magnetic Resonance (NMR) spectroscopy was employed.
  • Resonance assignments were obtained for (1)H, (15)N, and (13)C nuclei.
  • Data were deposited in the Biological Magnetic Resonance Data Bank (BMRB) under accession number 17288.

Main Results:

  • Complete (1)H, (15)N, and (13)C backbone and side-chain chemical shift assignments were achieved for Staphylococcus aureus MazF.
  • This represents the first reported solution NMR structure data for a wild-type MazF toxin.

Conclusions:

  • The provided chemical shift assignments are essential for determining the three-dimensional structure of Staphylococcus aureus MazF.
  • This work lays the groundwork for understanding the structure-function relationships of MazF toxins in Gram-positive pathogens.

More Related Videos

15N CPMG Relaxation Dispersion for the Investigation of Protein Conformational Dynamics on the µs-ms Timescale08:09

15N CPMG Relaxation Dispersion for the Investigation of Protein Conformational Dynamics on the µs-ms Timescale

Published on: April 19, 2021

MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria08:34

MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria

Published on: February 23, 2021

Related Concept Videos

NMR Spectroscopy: Chemical Shift Overview01:15

NMR Spectroscopy: Chemical Shift Overview

The position of the absorption signal of a sample is reported relative to the position of the signal of tetramethylsilane (TMS), which is added as an internal reference while recording spectra. The difference between the absorption frequencies of the sample and TMS (in Hz) is divided by the spectrometer operating frequency (in MHz) to obtain a dimensionless quantity called the chemical shift. It is reported on the δ (delta) scale and expressed in parts per million.
For instance, the proton...
Carbon-13 (¹³C) NMR: Overview01:10

Carbon-13 (¹³C) NMR: Overview

Carbon-13 is a naturally occurring NMR-active isotope of carbon with a low natural abundance of 1.1%. In contrast, carbon-12 is the most abundant isotope of carbon with zero nuclear spin. Therefore, it is NMR inactive. The gyromagnetic ratio of carbon-13 is smaller than that of protons. As a result, carbon-13 resonance is about 6000 times weaker than proton resonance. For a given magnetic field strength, the resonance frequency of carbon-13 is about one-fourth of the resonance frequency for...
¹H NMR: Complex Splitting01:13

¹H NMR: Complex Splitting

A proton M that is coupled to a proton X results in doublet signals for M. However, NMR-active nuclei can be simultaneously coupled to more than one nonequivalent nucleus. When M is coupled to a second proton A, such as in styrene oxide, each peak in the doublet is split into another doublet.
Splitting diagrams or splitting tree diagrams are routinely used to depict such complex couplings. While drawing splitting diagrams, the splitting with the larger coupling constant is usually applied first.
¹H NMR: Interpreting Distorted and Overlapping Signals01:02

¹H NMR: Interpreting Distorted and Overlapping Signals

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 slanted or...
¹H NMR of Conformationally Flexible Molecules: Temporal Resolution00:52

¹H NMR of Conformationally Flexible Molecules: Temporal Resolution

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...
Proton (¹H) NMR: Chemical Shift01:07

Proton (¹H) NMR: Chemical Shift

Organic molecules primarily contain carbon and hydrogen atoms. While all the hydrogen isotopes are NMR-active, protium or hydrogen-1 is the most abundant. It has a significant energy separation between its nuclear spin states due to its large gyromagnetic ratio. As per Boltzmann's distribution, an increase in the energy separation implies a greater excess population of nuclei available for excitation, resulting in a strong NMR absorption signal.
Absorption signals of all the protium nuclei in a...