Fast MAS Total Through-Bond Correlation Spectroscopy

Hardy, Edme H.; Verel, René; Meier, Beat H.

doi:10.1006/jmre.2000.2258

Cited by 105 publications

(89 citation statements)

References 31 publications

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“…MAS rates between 8 and 15 kHz were used at sample temperatures between Ϫ14°C and Ϫ5°C. For homonuclear through-bond polarization transfer, total through-bond-correlation spectroscopy (TOBSY) schemes were used (39)(40)(41). Selection of regions with high mobility was achieved by applying insensitive nuclei enhanced by polarization transfer (INEPT) magnetization transfer from protons to carbon atoms (42), and regions with intermediate mobility were selected by combining a proton T 2 -filter with CP magnetization transfer.…”

Section: Methodsmentioning

confidence: 99%

Molecular-level secondary structure, polymorphism, and dynamics of full-length α-synuclein fibrils studied by solid-state NMR

Heise

Hoyer

Becker

et al. 2005

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

611

728

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The 140-residue protein ␣-synuclein (AS) is able to form amyloid fibrils and as such is the main component of protein inclusions involved in Parkinson's disease. We have investigated the structure and dynamics of full-length AS fibrils by high-resolution solid-state NMR spectroscopy. Homonuclear and heteronuclear 2D and 3D spectra of fibrils grown from uniformly 13 C͞ 15 N-labeled AS and AS reverse-labeled for two of the most abundant amino acids, K and V, were analyzed. 13 C and 15 N signals exhibited linewidths of <0.7 ppm. Sequential assignments were obtained for 48 residues in the hydrophobic core region. We identified two different types of fibrils displaying chemical-shift differences of up to 13 ppm in the 15 N dimension and up to 5 ppm for backbone and side-chain 13 C chemical shifts. EM studies suggested that molecular structure is correlated with fibril morphology. Investigation of the secondary structure revealed that most amino acids of the core region belong to ␤-strands with similar torsion angles in both conformations. Selection of regions with different mobility indicated the existence of monomers in the sample and allowed the identification of mobile segments of the protein within the fibril in the presence of monomeric protein. At least 35 C-terminal residues were mobile and lacked a defined secondary structure, whereas the N terminus was rigid starting from residue 22. Our findings agree well with the overall picture obtained with other methods and provide insight into the amyloid fibril structure and dynamics with residue-specific resolution.EM ͉ protein structure ͉ amyloid ͉ Parkinson's disease ͉ protein aggregation T he ability of a protein to form amyloid fibrils is increasingly recognized as a general property of all polypeptide sequences (1). It is a phenomenon common to numerous neurodegenerative diseases such as Alzheimer's and Parkinson's diseases and spongiform encephalopathies (2, 3). Thus, the investigation of the mechanism(s) of protein misfolding as well as the detailed structures of amyloid fibrils is of paramount interest (4, 5). Parkinson's disease is defined by the presence of intracellular inclusions in dopaminergic neurons, the so-called Lewy bodies, which contain a high content of fibrils formed from the 140-aa cytoplasmic protein ␣-synuclein (AS) (6). The question of whether the mature fibrils themselves or rather protofilaments or folding intermediates are the neurotoxic species responsible for the cell death of dopaminergic neurons is still a subject of great controversy (7-9).AS belongs to the class of natively unfolded proteins, i.e., it lacks a well-defined secondary structure (10, 11), although long-range interactions have been shown to stabilize an aggregation-autoinhibited global protein architecture (12). Three regions of the protein can be classified. (i) The amphipathic N terminus (residues 1-60) consists of imperfect 11-mer repeats, with the consensus motif KTKEGV. (ii) The predominantly hydrophobic middle, also known as non-A␤ component region (residues 61-95...

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Section: Methodsmentioning

confidence: 99%

Molecular-level secondary structure, polymorphism, and dynamics of full-length α-synuclein fibrils studied by solid-state NMR

Heise

Hoyer

Becker

et al. 2005

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

611

728

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“…17,18 For improved spectral selectivity, however, it is useful to focus on the analysis of indirect (scalar) spin-spin interactions, which can discriminate through-bond connectivity from mere spatial proximity. Thus, a variety of two-dimensional total-through-bond correlation spectroscopic (TOBSY) approaches have been described in the literature, [19][20][21][22][23][24][25][26][27] enabling the selective correlation between those nuclei that are directly bonded. A particularly successful general concept for the development of suitable recoupling strategies has been the so-called RN n n pulse symmetry, introduced by Levitt and co-workers.…”

Section: Introductionmentioning

confidence: 99%

NMR studies of phosphorus chalcogenide–copper iodide coordination compounds

Brunklaus

Chan

Eckert

et al. 2003

Phys. Chem. Chem. Phys.

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The local structures of the new phosphorus chalcogenide -copper iodide coordination compounds (CuI)P 4 Se 4 , (CuI) 2 P 8 Se 3 , (CuI) 3 P 4 Se 4 , and (CuI) 3 P 4 S 4 are investigated using comprehensive 63 Cu, 65 Cu, and 31 P magic angle spinning NMR techniques. Peak assignments are proposed on the basis of homo-and heteronuclear indirect spin-spin interactions, available from lineshape analysis and/or two-dimensional correlation spectroscopy. In particular, the 31 P-63,65 Cu scalar coupling constants have been extracted from detailed lineshape simulations of the 31 P resonances associated with the Cu-bonded P atoms. In addition, the RN n n pulse symmetry concept of Levitt and coworkers has been utilized for total through-bond correlation spectroscopy (TOBSY) of directly-bonded phosphorus species. The resonance assignments obtained facilitate a discussion of the 31 P and 63,65 Cu NMR Hamiltonian parameters in terms of the detailed local atomic environments. Analysis of the limited data set available for this group of closely related compounds offers the following conclusions: (1) bonding of a special phosphorus site in a given P 4 X n (X ¼ S, Se) molecule to Cu + ions shifts the corresponding 31 P NMR signal upfield by about 50 ppm relative to the uncomplexed molecule, (2) the magnitude of the corresponding scalar 31 P-63,65 Cu spin-spin coupling constant tends to decrease with increasing Cu-P distance, and (3) the 63,65 Cu nuclear electric quadrupolar coupling constants appear to be weakly correlated with the shear strain parameter specifying the degree of local distortion present in the four-coordinated [CuI 2 P 2 ] and [CuI 3 P] environments. Overall, the results illustrate the power and potential of advanced solid state NMR methodology to provide useful structural information in this class of materials.

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“…To further assess the mobility, conformational heterogeneity, and secondary structure formation of MBP bound to actin, a series of two-dimensional 13 C-13 C correlation spectra were recorded using both through-bond (TOBSY (through-bond correlation spectroscopy); Hardy et al 2001) and through-space (DARR (dipolar-assisted rotational resonance; Takegoshi et al 2001) or SPC5 3 (Hohwy et al 2002)) interactions as mechanisms to establish molecular correlations. Through-space spectra acquired using either DARR mixing (50 ms) or the shorter mixing time of SPC5 3 (1.26 ms) supported the conclusions derived from the one-dimensional CP-MAS and INEPT-MAS spectra.…”

Section: Association With Actin Stabilizes Ordered Secondary Structurmentioning

confidence: 99%

Fuzzy complexes of myelin basic protein: NMR spectroscopic investigations of a polymorphic organizational linker of the central nervous systemThis paper is one of a selection of papers published in this special issue entitled “Canadian Society of Biochemistry, Molecular & Cellular Biology 52nd Annual Meeting — Protein Folding: Principles and Diseases” and has undergone the Journal's usual peer review process.

Libich

Ahmed

Zhong

et al. 2010

Biochem. Cell Biol.

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The classic 18.5 kDa isoform of myelin basic protein (MBP) is central to maintaining the structural homeostasis of the myelin sheath of the central nervous system. It is an intrinsically disordered, promiscuous, multifunctional, peripheral membrane protein, whose conformation adapts to its particular environment. Its study requires the selective and complementary application of diverse approaches, of which solution and solid-state NMR spectroscopy are the most powerful to elucidate site-specific features. We review here several recent solution and solid-state NMR spectroscopic studies of 18.5 kDa MBP, and the induced partial disorder-to-order transitions that it

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Fast MAS Total Through-Bond Correlation Spectroscopy

Cited by 105 publications

References 31 publications

Molecular-level secondary structure, polymorphism, and dynamics of full-length α-synuclein fibrils studied by solid-state NMR

Molecular-level secondary structure, polymorphism, and dynamics of full-length α-synuclein fibrils studied by solid-state NMR

NMR studies of phosphorus chalcogenide–copper iodide coordination compounds

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