%0 Journal Article %J Solid State Nuclear Magnetic Resonance %D 2006 %T Solid-state NMR structural studies of the fibril form of a mutant mouse prion peptide PrP89-143(P101L) %A Lim, K. H. %A Nguyen, T. N. %A Damo, S. M. %A Mazur, T. %A Ball, H. L. %A Prusiner, S. B. %A Pines, A. %A Wemmer, D. E. %K identification %X

The peptide fragment 89-143 of the prion protein (carrying a P101L mutation) is biologically active in transgenic mice when in a fibrillar form. Injection of these fibrils into transgenic mice (expressing full length PrP with the P101L mutation) induces a neurodegenerative prion disease (Kaneko et al., J. Mol. Biol. 295 (2000) 997). Here we present solid-state NMR studies of PrP89-143(P101L) fibrils, probing the conformation of residues in the hydrophobic segment 112-124 with chemical shifts. The conformations of glycine residues were analyzed using doubly C-13 = 0 labeled peptides by two-dimensional (2D) double-quantum correlation, and double-quantum filtered dephasing distance measurements. MQ-NMR experiments were carried out to probe the relative alignment of the individual peptides fibrils. These NMR studies indicate that the 112-124 segment adopts an extended beta-sheet conformation, though not in a parallel, in register alignment. There is evidence for conformational variability at Gly 113. DQ correlation experiments provide useful information in regions with conformational heterogeneity. (c) 2005 Elsevier Inc. All rights reserved.

%B Solid State Nuclear Magnetic Resonance %V 29 %P 183-190 %8 Feb %@ 0926-2040 %G English %U ://WOS:000234417500020 %N 1-3 %M WOS:000234417500020 %! Solid-state NMR structural studies of the fibril form of a mutant mouse prion peptide PrP89-143(P101L) %R Doi 10.1016/J.Ssnmr.2005.09.017 %0 Journal Article %J Proceedings of the National Academy of Sciences of the United States of America %D 2001 %T Solid-state NMR studies of the secondary structure of a mutant prion protein fragment of 55 residues that induces neurodegeneration %A Laws, D. D. %A Bitter, H. M. L. %A Liu, K. %A Ball, H. L. %A Kaneko, K. %A Wille, H. %A Cohen, F. E. %A Prusiner, S. B. %A Pines, A. %A Wemmer, D. E. %K diseases %X

The secondary structure of a 55-residue fragment of the mouse prion protein, MoPrP(89-143), was studied in randomly aggregated (dried from water) and fibrillar (precipitated from water/ acetonitrile) forms by C-13 solid-state NMR. Recent studies have shown that the fibrillar form of the P101L mutant of MoPrP(89-143) is capable of inducing prion disease in transgenic mice, whereas unaggregated or randomly aggregated samples do not provoke disease. Through analysis of C-13 chemical shifts, we have determined that both wild-type and mutant sequence MoPrP(89-143) form a mixture of beta -sheet and alpha -helical conformations in the randomly aggregated state although the beta -sheet content in MoPrP(89-143, P101L) is significantly higher than in the wild-type peptide. In a fibrillar state, MoPrP(89-143, P101L) is completely converted into beta -sheet, suggesting that the formation of a specific beta -sheet structure may be required for the peptide to induce disease. Studies of an analogous peptide from Syrian hamster PrP verify that sequence alterations in residues 101-117 affect the conformation of aggregated forms of the peptides.

%B Proceedings of the National Academy of Sciences of the United States of America %V 98 %P 11686-11690 %8 Sep 25 %@ 0027-8424 %G English %U ://WOS:000171237100125 %N 20 %M WOS:000171237100125 %! Solid-state NMR studies of the secondary structure of a mutant prion protein fragment of 55 residues that induces neurodegeneration %R Doi 10.1073/Pnas.201404298 %0 Journal Article %J Chemical Physics Letters %D 1996 %T Application of rotational resonance to inhomogeneously broadened systems %A Heller, J. %A Larsen, R. %A Ernst, M. %A Kolbert, A. C. %A Baldwin, M. %A Prusiner, S. B. %A Wemmer, D. E. %A Pines, A. %K nmr %X

A protocol is presented for the determination of internuclear distances using rotational-resonance magnetization-exchange NMR in systems with inhomogeneously broadened lines. Non-linear least-square fitting procedures are used to obtain the distance, the inhomogeneous broadening, the zero-quantum relaxation time, and error estimates for these parameters. We apply this procedure to a biological system of unknown structure.

%B Chemical Physics Letters %V 251 %P 223-229 %8 Mar 22 %@ 0009-2614 %G English %U ://WOS:A1996UD92400015 %N 3-4 %M WOS:A1996UD92400015 %! Application of rotational resonance to inhomogeneously broadened systems %R Doi 10.1016/0009-2614(96)00098-X %0 Journal Article %J Protein Science %D 1996 %T Solid-state NMR studies of the prion protein H1 fragment %A Heller, J. %A Kolbert, A. C. %A Larsen, R. %A Ernst, M. %A Bekker, T. %A Baldwin, M. %A Prusiner, S. B. %A Pines, A. %A Wemmer, D. E. %K alpha-helices %X

Conformational changes in the prion protein (PrP) seem to be responsible for prion diseases. We have used conformation-dependent chemical-shift measurements and rotational-resonance distance measurements to analyze the conformation of solid-state peptides lacking long-range order, corresponding to a region of PrP designated H1. This region is predicted to undergo a transformation of secondary structure in generating the infectious form of the protein. Solid-state NMR spectra of specifically C-13-enrrched samples of H1, residues 109-122 (MKHMAGAAAAGAVV) of Syrian hamster PrP, have been acquired under cross-polarization and magic-angle spinning conditions. Samples lyophilized from 50% acetonitrile/50% water show chemical shifts characteristic of a beta-sheet conformation in the region corresponding to residues 112-121, whereas samples lyophilized from hexafluoroisopropanol display shifts indicative of alpha-helical secondary structure in the region corresponding to residues 113-117. Complete conversion to the helical conformation was not observed and conversion from alpha-helix back to beta-sheet, as inferred from the solid-state NMR spectra, occurred when samples were exposed to water. Rotational-resonance experiments were performed on seven doubly C-13-labeled H1 samples dried from water. Measured distances suggest that the peptide is in an extended, possibly beta-strand, conformation. These results are consistent with the experimental observation that PrP can exist in different conformational states and with structural predictions based on biological data and theoretical modeling that suggest that H1 may play a key role in the conformational transition involved in the development of prion diseases.

%B Protein Science %V 5 %P 1655-1661 %8 Aug %@ 0961-8368 %G English %U ://WOS:A1996VA13800019 %N 8 %M WOS:A1996VA13800019 %! Solid-state NMR studies of the prion protein H1 fragment %0 Journal Article %J Journal of Physical Chemistry %D 1994 %T Measurement of Internuclear Distances by Switched Angle Spinning Nmr %A Kolbert, A. C. %A Grandinetti, P. J. %A Baldwin, M. %A Prusiner, S. B. %A Pines, A. %K protein %X

Dipolar switched angle spinning, an NMR technique due to Terao et al. [Terao, T.; Miura H.; Saika, A, J. Chem. Phys. 1986, 85, 3816], has been used to measure C-13-C-13 distances in the solid state. The experiment involves rotation of the sample at two different angles during different periods of a two-dimensional experiment. An evolution period with off-magic-angle spinning and chemical shift refocusing, followed by detection of the signal under magic angle spinning, yields scaled Pake patterns in omega(1), correlated with their isotropic shifts in omega(2) allowing the high-resolution measurement of dipolar couplings. We demonstrate this experiment on samples of doubly C-13 labeled zinc acetate and a 14 amino acid peptide, in which we show that under optimal conditions distances of up to 5 Angstrom may be measured.

%B Journal of Physical Chemistry %V 98 %P 7936-7938 %8 Aug 18 %@ 0022-3654 %G English %U ://WOS:A1994PC61900003 %N 33 %M WOS:A1994PC61900003 %! Measurement of Internuclear Distances by Switched Angle Spinning Nmr %R Doi 10.1021/J100084a003