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Related Concept Videos

Insensitive Nuclei Enhanced by Polarization Transfer (INEPT)01:15

Insensitive Nuclei Enhanced by Polarization Transfer (INEPT)

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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...
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Nuclear magnetic resonance (NMR) spectroscopy is a very valuable analytical technique for researchers. It has been used for more than 50 years as an analytical tool. F. Bloch and E. Purcell formulated NMR in 1946 and won the 1952 Nobel Prize in Physics  for their work. Biological macromolecules such as proteins, nucleic acids, lipids, and organic molecules including pharmaceutical compounds, can be studied using this versatile tool that exploits the magnetic properties of certain nuclei.
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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.
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The probability of having two carbon-13 atoms next to each other is negligible because of the low natural abundance of carbon-13. Consequently, peak splitting due to carbon-carbon spin-spin coupling is not observed in spectra. However, protons up to three sigma bonds away split the carbon signal according to the n+1 rule, resulting in complicated spectra.
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Related Experiment Video

Updated: Jun 3, 2025

Preparation of Fungal and Plant Materials for Structural Elucidation Using Dynamic Nuclear Polarization Solid-State NMR
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Preparation of Fungal and Plant Materials for Structural Elucidation Using Dynamic Nuclear Polarization Solid-State NMR

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Ultrasensitive Characterization of Native Bacterial Biofilms via Dynamic Nuclear Polarization-Enhanced Solid-State

Chang-Hyeock Byeon1, Ted Kinney1, Hakan Saricayir1

  • 1Department of Structural Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA, 15261, USA.

Angewandte Chemie (International Ed. in English)
|January 8, 2025
PubMed
Summary

Dynamic Nuclear Polarization (DNP) solid-state NMR (ssNMR) allows high-resolution structural analysis of native bacterial biofilms. This breakthrough enhances understanding of biofilm composition and structure, aiding in combating resistant infections.

Keywords:
MAS DNP ssNMRPseudomonas fluorescensbacterial colony biofilmbiofilm compositionhyperpolarized NMR

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Area of Science:

  • Microbiology
  • Biophysics
  • Biochemistry

Background:

  • Bacterial biofilms are crucial in persistent infections and antimicrobial resistance.
  • Analyzing native biofilm structure is challenging due to methodological limitations.
  • Solid-state NMR (ssNMR) offers potential but faces sensitivity issues with unlabeled samples.

Purpose of the Study:

  • To apply high-sensitivity Dynamic Nuclear Polarization (DNP) ssNMR for characterizing native bacterial biofilms.
  • To overcome sensitivity limitations of conventional ssNMR for biofilm analysis.
  • To enable structural characterization without isotope labeling or sample modification.

Main Methods:

  • Utilized DNP-enhanced ssNMR for structural characterization of native Pseudomonas fluorescens biofilms.
  • Acquired 1D and 2D ssNMR spectra (13C/15N, 1H-13C, 1H-15N, 13C-13C) rapidly.
  • Employed favorable freezing conditions for quantitative detection of flexible and rigid biofilm components.

Main Results:

  • Achieved ~75-fold sensitivity enhancement with DNP ssNMR.
  • Successfully identified and quantified extracellular matrix (ECM) components.
  • Enabled rapid data acquisition (seconds to hours) for biofilm analysis.

Conclusions:

  • This study is the first to use DNP ssNMR for native bacterial biofilm characterization.
  • DNP ssNMR significantly enhances capabilities for analyzing diverse in vitro and ex vivo biofilms.
  • This versatile approach will accelerate structure-guided strategies against biofilm-related infections.