Simultaneous acquisition of PAR and PAIN spectra

Nielsen, Anders B.; Székely, Kathrin; Gath, Julia; Ernst, Matthias; Nielsen, Niels Chr.; Meier, Beat H.

doi:10.1007/s10858-012-9616-7

Cited by 24 publications

(21 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This novel strategy allows the concatenation of 2D or 3D pulse sequences into a single experiment and thus potentially combines all the aforementioned advances, boosting the capacity of any NMR spectrometer at least two-fold without the need for sample manipulation or additional hardware. At the heart of this approach is simultaneous cross-polarization from 1 H to 13 C and 15 N, which has also been used in time-shared experiments (Linser et al 2011; Nielsen et al 2012), and long lived 15 N longitudinal magnetization (Giraud et al 2005) which can be stored and recalled to perform additional experiments. The DUMAS scheme enables one to acquire two experiments: the first with 100% of the sensitivity with respect to classical pulse sequences, and a second experiment generated from the 15 N polarization with typically 80% of the sensitivity (Gopinath and Veglia 2012a).…”

Section: Introductionmentioning

confidence: 99%

Determination of structural topology of a membrane protein in lipid bilayers using polarization optimized experiments (POE) for static and MAS solid state NMR spectroscopy

2013

View full text Add to dashboard Cite

The low sensitivity inherent to both the static and magic angle spinning techniques of solid-state NMR (ssNMR) spectroscopy has thus far limited the routine application of multidimensional experiments to determine the structure of membrane proteins in lipid bilayers. Here, we demonstrate the advantage of using a recently developed class of experiments, polarization optimized experiments (POE), for both static and MAS spectroscopy to achieve higher sensitivity and substantial time-savings for 2D and 3D experiments. We used sarcolipin, a single pass membrane protein, reconstituted in oriented bicelles (for oriented ssNMR) and multilamellar vesicles (for MAS ssNMR) as a benchmark. The restraints derived by these experiments are then combined into a hybrid energy function to allow simultaneous determination of structure and topology. The resulting structural ensemble converged to a helical conformation with a backbone RMSD ∼ 0.44 Å, a tilt angle of 24° ± 1°, and an azimuthal angle of 55° ± 6°. This work represents a crucial first step toward obtaining high-resolution structures of large membrane proteins using combined multidimensional O-ssNMR and MAS-ssNMR.

show abstract

Section: Introductionmentioning

confidence: 99%

Determination of structural topology of a membrane protein in lipid bilayers using polarization optimized experiments (POE) for static and MAS solid state NMR spectroscopy

2013

View full text Add to dashboard Cite

show abstract

“…It now includes the 1 H detected experiments under fast MAS conditions [46], the afterglow sequence developed by Traaseth and coworkers [47, 48], the simultaneous acquisition of PAIN and PAR experiments discovered by Meier and coworkers [49] as well as the more recent pulse sequences that are the dipolar-edited versions of DUMAS methodology [50]. These experiments reinforce the importance of developing pulse sequences that exploit as many orphan operators as possible to enhance the NMR signal in solids and enable multiple acquisitions.…”

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

View full text Add to dashboard Cite

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.

show abstract

“…It is to be noted that at higher spinning speeds (∼20 kHz) the sensitivity of 13 C and 15 N SIM-CP is almost identical to that of double resonance CP. 34 For specific-CP from 15 N to 13 Cα (or 13 C ), the 13 C offset was shifted to 60 (or 177) ppm. During specific-CP, the 15 N RF amplitude was set to (5/2) · ω r = 32.5 kHz, whereas 13 C RF amplitude was set to (3/2) · ω r (=19.5 kHz) and (7/2) · ω r (=45.5 kHz) for 13 Cα and 13 C transfers, respectively.…”

Section: Methodsmentioning

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

View full text Add to dashboard Cite

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.

show abstract

Simultaneous acquisition of PAR and PAIN spectra

Cited by 24 publications

References 40 publications

Determination of structural topology of a membrane protein in lipid bilayers using polarization optimized experiments (POE) for static and MAS solid state NMR spectroscopy

Determination of structural topology of a membrane protein in lipid bilayers using polarization optimized experiments (POE) for static and MAS solid state NMR spectroscopy

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

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

Contact Info

Product

Resources

About