Dual Acquisition Magic‐Angle Spinning Solid‐State NMR‐Spectroscopy: Simultaneous Acquisition of Multidimensional Spectra of Biomacromolecules

Gopinath, T.; Veglia, Gianluigi

doi:10.1002/anie.201108132

Cited by 78 publications

(99 citation statements)

References 59 publications

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“…The sequence for a three-dimensional 1 H detection experiment is shown in Figure 1A. One and two-dimensional experiments with 1 H detection and simultaneous correlation to 13 C and 15 N nuclei were adapted from previous publications using simultaneous cross polarization (CP) [14, 15, 17] and concurrent cross polarization pulse schemes [27]. 1 H magnetization is transferred to 13 C and 15 N simultaneously using Hartmann-Hahn CP.…”

Section: Resultsmentioning

confidence: 99%

“…Notably, 13 C/ 13 C homonuclear correlation under proton assisted recoupling (PAR) and 13 C/ 15 N heteronuclear correlation under proton-assisted insensitive nuclei (PAIN) can be performed using simultaneous spin-lock pulses on three channels under second-order recoupling schemes. In addition, recently, it has been shown that bidirectional cross-polarization between 13 C and 15 N nuclei can be used to transfer coherence without significant losses in signal intensities [15, 17]. This has enabled triple-resonance experiments to be executed with multiple acquisitions using a single receiver within a single experiment.…”

Section: Introductionmentioning

confidence: 99%

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Simultaneous cross polarization to 13C and 15N with 1H detection at 60 kHz MAS solid-state NMR

Das

Opella

2016

Journal of Magnetic Resonance

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We describe high resolution MAS solid-state NMR experiments that utilize 1H detection with 60 kHz magic angle spinning; simultaneous cross-polarization from 1H to 15N and 13C nuclei; bidirectional cross-polarization between 13C and 15N nuclei; detection of both amide nitrogen and aliphatic carbon 1H; and measurement of both 13C and 15N chemical shifts through multi-dimensional correlation experiments. Three-dimensional experiments correlate amide 1H and alpha 1H selectively with 13C or 15N nuclei in a polypeptide chain. Two separate three-dimensional spectra correlating 1Hα/13Cα/1HN and 1HN/15N/1Hα are recorded simultaneously in a single experiment, demonstrating that a two-fold savings in experimental time is potentially achievable. Spectral editing using bidirectional coherence transfer pathways enables simultaneous magnetization transfers between 15N, 13Cα(i) and 13C′(i−1), facilitating intra- and inter- residue correlations for sequential resonance assignment. Non-uniform sampling is integrated into the experiments, further reducing the length of experimental time.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Simultaneous cross polarization to 13C and 15N with 1H detection at 60 kHz MAS solid-state NMR

Das

Opella

2016

Journal of Magnetic Resonance

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“…In recent years, our group has been redesigning the classical MAS ssNMR experiments for resonance assignment and distance determination in biological solids, boosting both sensitivity and resolution [9–12]. Toward this goal, we developed a new class of experiments called Polarization Optimized Experiments (POE) that utilize orphan spin operators to generate up to eight multi-dimensional NMR spectra from one pulse sequence [9, 13].…”

Section: Introductionmentioning

confidence: 99%

“…Toward this goal, we developed a new class of experiments called Polarization Optimized Experiments (POE) that utilize orphan spin operators to generate up to eight multi-dimensional NMR spectra from one pulse sequence [9, 13]. These experiments are performed using commercial ssNMR probes for bio-solids and require only one receiver.…”

Section: Introductionmentioning

confidence: 99%

1 H-detected MAS solid-state NMR experiments enable the simultaneous mapping of rigid and dynamic domains of membrane proteins

Gopinath

Nelson

Veglia

2017

Journal of Magnetic Resonance

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Magic angle spinning (MAS) solid-state NMR (ssNMR) spectroscopy is emerging as a unique method for the atomic resolution structure determination of native membrane proteins in lipid bilayers. Although 13C-detected ssNMR experiments continue to play a major role, recent technological developments have made it possible to carry out 1H-detected experiments, boosting both sensitivity and resolution. Here, we describe a new set of 1H-detected hybrid pulse sequences that combine through-bond and through-space correlation elements into single experiments, enabling the simultaneous detection of rigid and dynamic domains of membrane proteins. As proof-of-principle, we applied these new pulse sequences to the membrane protein phospholamban (PLN) reconstituted in lipid bilayers under moderate MAS conditions. The cross-polarization (CP) based elements enabled the detection of the relatively immobile residues of PLN in the transmembrane domain using through-space correlations; whereas the most dynamic region, which is in equilibrium between folded and unfolded states, was mapped by through-bond INEPT-based elements. These new 1H-detected experiments will enable one to detect not only the most populated (ground) states of biomacromolecules, but also sparsely populated high-energy (excited) states for a complete characterization of protein free energy landscapes.

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“…While in liquid-state NMR there are several pulse sequences designed to detect discarded coherences via dual receivers or phase cycling schemes, 4 in ssNMR there are only a few examples that utilize orphan (i.e., neglected) spin operators to boost sensitivity. [5][6][7][8][9][10][11][12][13] In general, classical ssNMR pulse sequences eliminate orphan spin operators of magnetization either using z-filters or phase cycling schemes that suppress the spectral artifacts arising from undesired coherence transfer pathways.…”

Section: Introductionmentioning

confidence: 99%

Orphan spin operators enable the acquisition of multiple 2D and 3D magic angle spinning solid-state NMR spectra

Gopinath

Veglia

2013

The Journal of Chemical Physics

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We propose a general method that enables the acquisition of multiple 2D and 3D solid-state NMR spectra for U-13 C, 15 N-labeled proteins. This method, called MEIOSIS (Multiple ExperIments via Orphan SpIn operatorS), makes it possible to detect four coherence transfer pathways simultaneously, utilizing orphan (i.e., neglected) spin operators of nuclear spin polarization generated during 15 N-13 C cross polarization (CP). In the MEIOSIS experiments, two phase-encoded free-induction decays are decoded into independent nuclear polarization pathways using Hadamard transformations. As a proof of principle, we show the acquisition of multiple 2D and 3D spectra of U-13 C, 15 N-labeled microcrystalline ubiquitin. Hadamard decoding of CP coherences into multiple independent spin operators is a new concept in solid-state NMR and is extendable to many other multidimensional experiments. The MEIOSIS method will increase the throughput of solid-state NMR techniques for microcrystalline proteins, membrane proteins, and protein fibrils. © 2013 AIP Publishing LLC.

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Dual Acquisition Magic‐Angle Spinning Solid‐State NMR‐Spectroscopy: Simultaneous Acquisition of Multidimensional Spectra of Biomacromolecules

Cited by 78 publications

References 59 publications

Simultaneous cross polarization to 13C and 15N with 1H detection at 60 kHz MAS solid-state NMR

Simultaneous cross polarization to 13C and 15N with 1H detection at 60 kHz MAS solid-state NMR

1 H-detected MAS solid-state NMR experiments enable the simultaneous mapping of rigid and dynamic domains of membrane proteins

Orphan spin operators enable the acquisition of multiple 2D and 3D magic angle spinning solid-state NMR spectra

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