Selective averaging for high-resolution solid-state NMR spectroscopy of aligned samples

Nevzorov, Alexander A.; Opella, Stanley J.

doi:10.1016/j.jmr.2006.09.006

Cited by 142 publications

(151 citation statements)

References 44 publications

(64 reference statements)

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“…Solid-state NMR experiments PISEMA (20) and SAMPI4 (21) were acquired at 30°C. Spectra of [U- 15 N] samples were acquired with 1 k scans and 30-t1 increments, whereas selectively labeled samples required 4-to 12 k scans and Ϸ8 -16 increments.…”

Section: Nmr Spectroscopymentioning

confidence: 99%

Structure and topology of monomeric phospholamban in lipid membranes determined by a hybrid solution and solid-state NMR approach

Traaseth

Shi²,

Verardi

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

157

194

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Phospholamban (PLN) is an essential regulator of cardiac muscle contractility. The homopentameric assembly of PLN is the reservoir for active monomers that, upon deoligomerization form 1:1 complexes with the sarco(endo)plasmic reticulum Ca 2؉ -ATPase (SERCA), thus modulating the rate of calcium uptake. In lipid bilayers and micelles, monomeric PLN exists in equilibrium between a bent (or resting) T state and a more dynamic (or active) R state. Here, we report the high-resolution structure and topology of the T state of a monomeric PLN mutant in lipid bilayers, using a hybrid of solution and solid-state NMR restraints together with molecular dynamics simulations in explicit lipid environments. Unlike the previous structural ensemble determined in micelles, this approach gives a complete picture of the PLN monomer structure in a lipid bilayer. This hybrid ensemble exemplifies the tilt, rotation, and depth of membrane insertion, revealing the interaction with the lipids for all protein domains. The N-terminal amphipathic helical domain Ia (residues 1-16) rests on the surface of the lipid membrane with the hydrophobic face of domain Ia embedded in the membrane bilayer interior. The helix comprised of domain Ib (residues 23-30) and transmembrane domain II (residues 31-52) traverses the bilayer with a tilt angle of Ϸ24°. The specific interactions between PLN and lipid membranes may represent an additional regulatory element of its inhibitory function. We propose this hybrid method for the simultaneous determination of structure and topology for membrane proteins with compact folds or proteins whose spatial arrangement is dictated by their specific interactions with lipid bilayers.hybrid method ͉ membrane proteins ͉ oriented solid-state NMR ͉ molecular modeling ͉ PISEMA S tructure and topology are central to membrane protein function (1). Recently determined high-resolution structures reveal the compact folds for several membrane proteins, such as electron and proton-conducting proteins involved in photosynthesis and respiration (http://blanco.biomol.uci.edu/ Membrane Proteins xtal.html). However, a significant population of membrane proteins does not possess a compact tertiary fold, but has its fold space defined through interactions of secondary structure elements (helices, turns, and loops) with the lipid membrane, i.e., the topology (1). This is the case for phospholamban (PLN), a mammalian protein that is essential in the regulation of cardiac muscle contractility (2), and that has recently become a major target for gene therapy to ameliorate cardiomyopathies (3, 4). PLN is located in the sarco(endo)plasmic reticulum (SR) of cardiac myocytes, inhibiting the SR Ca 2ϩ -ATPase (SERCA) by shifting its relative Ca 2ϩ affinity (5). In vitro and in vivo experiments have shown PLN to exist as a homopentamer that deoligomerizes into active monomers that bind SERCA in a 1:1 molar ratio (6). The monomeric form of PLN exists in equilibrium between a dynamically disordered R state and a more restricted T state (7,8) and has 3 ...

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Section: Nmr Spectroscopymentioning

confidence: 99%

Structure and topology of monomeric phospholamban in lipid membranes determined by a hybrid solution and solid-state NMR approach

Traaseth

Shi²,

Verardi

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

157

194

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“…10 SLF experiments have been widely used by chemists and biophysicists to characterize the structures of liquid crystals as well as membrane proteins in mechanically or magnetically aligned lipid bilayers. [11][12][13][14][15][16][17][18] Since the introduction of the SLF experiment, several variants have emerged, such as PISEMA, [19][20][21] SAMPI4, 22 HIMSELF, 23 and more recently the sensitivity-enhanced and constant time versions. [24][25][26][27] These experiments are also known as rotating frame SLF (R-SLF) experiments and take advantage of the heteronuclear DC evolution under Hartmann-Hahn matching of RF fields for I and S spins and homonuclear decoupling for the abundant I spins.…”

Section: Introductionmentioning

confidence: 99%

Proton evolved local field solid-state nuclear magnetic resonance using Hadamard encoding: Theory and application to membrane proteins

Gopinath¹,

Mote²,

Veglia

2011

The Journal of Chemical Physics

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NMR anisotropic parameters such as dipolar couplings and chemical shifts are central to structure and orientation determination of aligned membrane proteins and liquid crystals. Among the separated local field experiments, the proton evolved local field (PELF) scheme is particularly suitable to measure dynamically averaged dipolar couplings and give information on local molecular motions. However, the PELF experiment requires the acquisition of several 2D datasets at different mixing times to optimize the sensitivity for the complete range of dipolar couplings of the resonances in the spectrum. Here, we propose a new PELF experiment that takes the advantage of the Hadamard encoding (HE) to obtain higher sensitivity for a broad range of dipolar couplings using a single 2D experiment. The HE scheme is obtained by selecting the spin operators with phase switching of hard pulses. This approach enables one to detect four spin operators, simultaneously, which can be processed into two 2D spectra covering a broader range of dipolar couplings. The advantages of the new approach are illustrated for a U-15 N NAL single crystal and the U-15 N labeled single-pass membrane protein sarcolipin reconstituted in oriented lipid bicelles. The HE-PELF scheme can be implemented in other multidimensional experiments to speed up the characterization of the structure and dynamics of oriented membrane proteins and liquid crystalline samples.

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“…Low sensitivity of NMR spectrometers greatly limited a use of the Pake powder patterns, and many other less direct NMR techniques for obtaining information about dipolar couplings have been developed. SLF (separated-local-field) spectroscopy (3), PISEMA (polarization inversion spin exchange at magic angle) (4,5), as well as more recent techniques: HIMSELF (heteronuclear isotropic mixing leading to spin exchange via the local field) (6), PIT-ANSEMA (polarization inversion time averaged nutation spin exchange at the magic angle) (7), or SAMMY (magic sandwiches) (8) have been used to measure distances between rare and abundant nuclei in oriented samples and single crystals when information about orientation is known. Higher efficiency of heteronuclear decoupling, compared to homonuclear, suggests that better accuracy can be achieved for well-isolated pairs of rare nuclei when the abundant nuclei (protons) are decoupled.…”

Section: Introductionmentioning

confidence: 99%

High‐precision measurement of internuclear distances using solid‐state NMR

Lee

Khitrin

2008

Concepts Magnetic Resonance

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Today, nuclear magnetic resonance (NMR) is among the most efficient tools in structural studies. Measurement of interatomic distances is the most common way of determining high-resolution structures of molecules using NMR techniques. In this article, we describe NMR techniques for static powder samples, based on a two-dimensional single-echo scheme, enhanced with adiabatic cross-polarization. They can significantly increase the accuracy of measuring internuclear distances and turn NMR into a high-precision crystallographic technique, complementing the X-ray, and neutron-scattering methods. Experimental examples are presented for intramolecular CÀ ÀN and CÀ ÀC distances in a-crystalline form of glycine.

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Selective averaging for high-resolution solid-state NMR spectroscopy of aligned samples

Cited by 142 publications

References 44 publications

Structure and topology of monomeric phospholamban in lipid membranes determined by a hybrid solution and solid-state NMR approach

Structure and topology of monomeric phospholamban in lipid membranes determined by a hybrid solution and solid-state NMR approach

Proton evolved local field solid-state nuclear magnetic resonance using Hadamard encoding: Theory and application to membrane proteins

High‐precision measurement of internuclear distances using solid‐state NMR

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