Methods for acquisition and assignment of multidimensional high-resolution magic angle spinning NMR of whole cell bacteria

Wei Li, Robin E.B. Lee, Richard E. Lee, Jinghu Li

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

High-resolution magic-angle spinning (HR-MAS) NMR was developed in late 1990s, and it has evolved quickly for the study of a variety of biological matrixes. Recently, it has been used as an effective means to study the cell wall structures of intact bacteria. 1H-13C heteronuclear single quantum coherence (HSQC) HR-MAS NMR can provide rapid analysis of the cell wall structure in live bacterial cells, thus allowing observation of drug effects, gene mutation, species differentiation, and environmental effects. However, this rapid analysis is dependent on having an established framework of HR-MAS NMR experiments and a detailed assignment of the whole-cell NMR spectra. This study examines parameters and describes strategies for the effective application of 2D and 3D HR-MAS NMR techniques to assign and study bacterial cell wall structures using Mycobacterium smegmatis as a model organism. Important parameters for successful whole-cell HR-MAS NMR studies, including pulse sequences, rotor synchronization, acquisition times, labeling strategies, temperature, number of cells, and cell viability, are described. A four-prong approach is presented for assignment of the complex whole-cell spectra, including the use of 3D HCCH-TOCSY and HCCH-COSY HR-MAS NMR.

Original languageEnglish (US)
Pages (from-to)5785-5792
Number of pages8
JournalAnalytical Chemistry
Volume77
Issue number18
DOIs
StatePublished - Sep 15 2005

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Magic angle spinning
Bacteria
Nuclear magnetic resonance
Cells
Labeling
Environmental impact
Synchronization
Rotors
Genes
Pharmaceutical Preparations

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

Cite this

Methods for acquisition and assignment of multidimensional high-resolution magic angle spinning NMR of whole cell bacteria. / Li, Wei; Lee, Robin E.B.; Lee, Richard E.; Li, Jinghu.

In: Analytical Chemistry, Vol. 77, No. 18, 15.09.2005, p. 5785-5792.

Research output: Contribution to journalArticle

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