Simultaneous acquisition of 13Cα–15N and 1H–15N–15N sequential correlations in proteins: application of dual receivers in 3D HNN

Chakraborty, Swagata; Paul, Subhradip; Hosur, Ramakrishna V.

doi:10.1007/s10858-011-9596-z

Cited by 28 publications

(21 citation statements)

References 12 publications

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“…[4][5][6][7][8][9][10][11] Following the introduction of parallel NMR spectroscopy, it was shown that one of its main advantages results from its use to collect two or more different kinds of multidimensional correlation experiments, within the time duration that would normally entail to collect a single spectrum. [5][6][7] These "parallel acquisition NMR, all-in-one combination of experimental applications" (PANACEA) strategies, have since been extended to systems of various heteronuclei [8,9] and adapted to protein liquid-state [10,11] and solid-state NMR experiments. [5][6][7] These "parallel acquisition NMR, all-in-one combination of experimental applications" (PANACEA) strategies, have since been extended to systems of various heteronuclei [8,9] and adapted to protein liquid-state [10,11] and solid-state NMR experiments.…”

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Multiple Parallel 2D NMR Acquisitions in a Single Scan

2013

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Multiple Parallel 2D NMR Acquisitions in a Single Scan

2013

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“…More recently, the afterglow phenomenon exploited by Kay and co-workers opened up the possibility for the acquisition of multidimensional NMR spectra of proteins in parallel [39]. The latter approach has been implemented with either one or multiple receivers [40-42]. Sensitivity enhanced experiments in solids have also been proposed by Tycko and co-workers [43] and been implemented in a manner similar to the sensitivity enhancement for solution NMR experiments.…”

Section: Discussionmentioning

confidence: 99%

Multiple acquisition of magic angle spinning solid-state NMR experiments using one receiver: Application to microcrystalline and membrane protein preparations

Gopinath

Veglia

2015

Journal of Magnetic Resonance

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Solid-State NMR spectroscopy of proteins is a notoriously low-throughput technique. Relatively low-sensitivity and poor resolution of protein samples require long acquisition times for multidimensional NMR experiments. To speed up data acquisition, we developed a family of experiments called Polarization Optimized Experiments (POE), in which we utilized the orphan spin operators that are discarded in classical multidimensional NMR experiments, recovering them to allow simultaneous acquisition of multiple 2D and 3D experiments, all while using conventional probes with spectrometers equipped with one receiver. POEs allow the concatenation of multiple 2D or 3D pulse sequences into a single experiment, thus potentially combining all of the aforementioned advances, boosting the capability of ssNMR spectrometers at least two-fold without the addition of any hardware. In this Perspective, we describe the first generation of POEs, such as dual acquisition MAS (or DUMAS) methods, and then illustrate the evolution of these experiments into MEIOSIS, a method that enables the simultaneous acquisition of multiple 2D and 3D spectra. Using these new pulse schemes for the solid-state NMR investigation of biopolymers makes it possible to obtain sequential resonance assignments, as well as distance restraints, in about half the experimental time. While designed for acquisition of heteronuclei, these new experiments can be easily implemented for proton detection and coupled with other recent advancements, such as dynamic polarization, to improve signal to noise. Finally, we illustrate the application of these methods to microcrystalline protein preparations as well as single and multi-span membrane proteins reconstituted in lipid membranes.

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“…It is worth pointing out that, unlike the INEPT procedure, the application of band‐selective 15 N‐ 13 C α HEHAHA mixing sequence for transferring the 15 N i magnetisation to the adjacent “ i +1” and “ i −1” residues makes it possible to conveniently extract the side chain chemical shift information simultaneously. The pulse sequences can also be adapted, using dual receivers as in earlier studies to acquire simultaneously RN[N‐CA HEHAHA]NCO and RN[N‐CA HEHAHA]CAHA correlation data sets. Another approach to simultaneously extract the side chain chemical shifts, using only a single data acquisition, would be to implement RF pulse schemes starting with the excitation of side chain protons such as the HSCSCACON[N‐CA HEHAHA]NH experiment involving the 1 H sc → 13 C sc → 13 C α magnetisation transfer pathway and chemical shift labelling of the side chain resonances in the indirect dimensions (see SI).…”

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confidence: 99%

A Set of Efficient nD NMR Protocols for Resonance Assignments of Intrinsically Disordered Proteins

et al. 2016

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The RF pulse scheme RN[N-CA HEHAHA]NH, which provides a convenient approach to the acquisition of different multidimensional chemical shift correlation NMR spectra leading to backbone resonance assignments, including those of the proline residues of intrinsically disordered proteins (IDPs), is experimentally demonstrated. Depending on the type of correlation data required, the method involves the generation of in-phase ((15) N)(x) magnetisation via different magnetisation transfer pathways such as H→N→CO→N, HA→CA→CO→N, H→N→CA→N and H→CA→N, the subsequent application of (15) N-(13) C(α) heteronuclear Hartmann-Hahn mixing over a period of ≈100 ms, chemical-shift labelling of relevant nuclei before and after the heteronuclear mixing step and amide proton detection in the acquisition dimension. It makes use of the favourable relaxation properties of IDPs and the presence of (1) JCαN and (2) JCαN couplings to achieve efficient correlation of the backbone resonances of each amino acid residue "i" with the backbone amide resonances of residues "i-1" and "i+1". It can be implemented in a straightforward way through simple modifications of the RF pulse schemes commonly employed in protein NMR studies. The efficacy of the approach is demonstrated using a uniformly ((15) N,(13) C) labelled sample of α-synuclein. The different possibilities for obtaining the amino-acid-type information, simultaneously with the connectivity data between the backbone resonances of sequentially neighbouring residues, have also been outlined.

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Simultaneous acquisition of 13Cα–15N and 1H–15N–15N sequential correlations in proteins: application of dual receivers in 3D HNN

Cited by 28 publications

References 12 publications

Multiple Parallel 2D NMR Acquisitions in a Single Scan

Multiple Parallel 2D NMR Acquisitions in a Single Scan

Multiple acquisition of magic angle spinning solid-state NMR experiments using one receiver: Application to microcrystalline and membrane protein preparations

A Set of Efficient nD NMR Protocols for Resonance Assignments of Intrinsically Disordered Proteins

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