In-Cell Solid-State NMR: An Emerging Technique for the Study of Biological Membranes

Warnet, Xavier L.; Arnold, Alexandre A.; Marcotte, Isabelle; Warschawski, Dror E.

doi:10.1016/j.bpj.2015.10.041

Cited by 46 publications

(32 citation statements)

References 64 publications

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“…In-cell solid-state NMR is an emerging field that requires new tools to expand its range of applicability, and 31 P NMR is underrepresented as one of these tools (44,45). Recently, 2 H NMR has been adapted to in-cell and even in vivo solid-state NMR to characterize lipid acyl chains in intact bacterial membranes and examine the effect of antimicrobial peptides (6,7).…”

Section: Discussionmentioning

confidence: 99%

A New Method of Assessing Lipid Mixtures by 31P Magic-Angle Spinning NMR

Warschawski

Arnold

Marcotte

2018

Biophysical Journal

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A variety of lipids that differ by their chains and headgroups are found in biomembranes. In addition to studying the overall membrane phase, determination of the structure, dynamics, and headgroup conformation of individual lipids in the mixture would be of great interest. We have thus developed, to our knowledge, a new approach using solid-state P NMR, magic-angle spinning, and chemical-shift anisotropy (CSA) recoupling, using an altered version of the recoupling of chemical shift anisotropy (ROCSA) pulse sequence, here penned PROCSA. The resulting two-dimensional spectra allowed the simultaneous measurement of the isotropic chemical shift and CSA of each lipid headgroup, thus providing a valuable measure of its dynamics and structure. PROCSA was applied to mixtures of phosphatidylethanolamine (PE) and phosphatidylglycerol (PG) in various relative proportions, to mimic bacterial membranes and assess the respective roles of lipids in shaping these bilayers. The results were interpreted in terms of membrane topology, lipid propensity to adopt various phases or conformations, and lipid-lipid miscibility. Our results showed that PG dictates the lipid behavior when present in a proportion of 20 mol % or more. A small proportion of PG is thus able to impose a bilayer structure to the hexagonal phase forming PE. We discuss the requirement for lipids, such as PE, to be able to adopt non-bilayer phases in a membrane.

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Section: Discussionmentioning

confidence: 99%

A New Method of Assessing Lipid Mixtures by 31P Magic-Angle Spinning NMR

Warschawski

Arnold

Marcotte

2018

Biophysical Journal

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“…Specific interactions, even transient interactions, with other cellular components will slow tumbling, resulting in broad resonances that are beyond detection. Solid-state NMR is not limited by molecular tumbling rate and therefore may be a better option for studying large-cell surface PSs (91,92). By using the high-resolution magic-angle spinning (HR-MAS) technique, Jachymek et al (93) demonstrated that the immunodominant O-polysaccharide on bacterial surfaces had the same structure as that in lipopolysaccharides and in isolated PSs.…”

Section: Magnetic Resonance Studies Of Cell Surface Carbohydratesmentioning

confidence: 99%

Magnetic Resonance Spectroscopy as a Tool for Assessing Macromolecular Structure and Function in Living Cells

Jie

Cheng

et al. 2017

Annual Rev. Anal. Chem.

214

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Investigating the structure, modification, interaction, and function of biomolecules in their native cellular environment leads to physiologically relevant knowledge about their mechanisms, which will benefit drug discovery and design. In recent years, nuclear and electron magnetic resonance (NMR) spectroscopy has emerged as a useful tool for elucidating the structure and function of biomacromolecules, including proteins, nucleic acids, and carbohydrates in living cells at atomic resolution. In this review, we summarize the progress and future of in-cell NMR as it is applied to proteins, nucleic acids, and carbohydrates.

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“…In-cell solution NMR spectroscopy has already been well established [114–117], with notable applications including the maturation of human superoxide dismutase [118,119], in cell characterization of alpha synuclein [120], and investigations of the relationship between molecular crowding and protein interactions [121,122]. In-cell solid state NMR [123], while a much less mature field, has been demonstrated to have promise on systems such as membrane proteins [124] or proteins in large complexes [125]. Many proteins of interest are dilute in cell, and have concentrations below the limit of detection for traditional solid-state NMR methods.…”

Section: Dnp In Cell Applications: Challenges and Opportunitiesmentioning

confidence: 99%

New NMR tools for protein structure and function: Spin tags for dynamic nuclear polarization solid state NMR

Rogawski

McDermott

2017

Archives of Biochemistry and Biophysics

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Magic angle spinning solid state NMR studies of biological macromolecules [1–3] have enabled exciting studies of membrane proteins [4,5], amyloid fibrils [6], viruses, and large macromolecular assemblies [7]. Dynamic nuclear polarization (DNP) provides a means to enhance detection sensitivity for NMR, particularly for solid state NMR, with many recent biological applications and considerable contemporary efforts towards elaboration and optimization of the DNP experiment. This review explores precedents and innovations in biological DNP experiments, especially highlighting novel chemical biology approaches to introduce the radicals that serve as a source of polarization in DNP experiments.

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In-Cell Solid-State NMR: An Emerging Technique for the Study of Biological Membranes

Cited by 46 publications

References 64 publications

A New Method of Assessing Lipid Mixtures by 31P Magic-Angle Spinning NMR

A New Method of Assessing Lipid Mixtures by 31P Magic-Angle Spinning NMR

Magnetic Resonance Spectroscopy as a Tool for Assessing Macromolecular Structure and Function in Living Cells

New NMR tools for protein structure and function: Spin tags for dynamic nuclear polarization solid state NMR

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