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. 1996 Aug;5(8):1655–1661. doi: 10.1002/pro.5560050819

Solid-state NMR studies of the prion protein H1 fragment.

J Heller 1, A C Kolbert 1, R Larsen 1, M Ernst 1, T Bekker 1, M Baldwin 1, S B Prusiner 1, A Pines 1, D E Wemmer 1
PMCID: PMC2143492  PMID: 8844854

Abstract

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 13C-enriched 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 13C-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.

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Selected References

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  1. Borchelt D. R., Scott M., Taraboulos A., Stahl N., Prusiner S. B. Scrapie and cellular prion proteins differ in their kinetics of synthesis and topology in cultured cells. J Cell Biol. 1990 Mar;110(3):743–752. doi: 10.1083/jcb.110.3.743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Büeler H., Aguzzi A., Sailer A., Greiner R. A., Autenried P., Aguet M., Weissmann C. Mice devoid of PrP are resistant to scrapie. Cell. 1993 Jul 2;73(7):1339–1347. doi: 10.1016/0092-8674(93)90360-3. [DOI] [PubMed] [Google Scholar]
  3. Büeler H., Fischer M., Lang Y., Bluethmann H., Lipp H. P., DeArmond S. J., Prusiner S. B., Aguet M., Weissmann C. Normal development and behaviour of mice lacking the neuronal cell-surface PrP protein. Nature. 1992 Apr 16;356(6370):577–582. doi: 10.1038/356577a0. [DOI] [PubMed] [Google Scholar]
  4. Caughey B. W., Dong A., Bhat K. S., Ernst D., Hayes S. F., Caughey W. S. Secondary structure analysis of the scrapie-associated protein PrP 27-30 in water by infrared spectroscopy. Biochemistry. 1991 Aug 6;30(31):7672–7680. doi: 10.1021/bi00245a003. [DOI] [PubMed] [Google Scholar]
  5. Cohen F. E., Pan K. M., Huang Z., Baldwin M., Fletterick R. J., Prusiner S. B. Structural clues to prion replication. Science. 1994 Apr 22;264(5158):530–531. doi: 10.1126/science.7909169. [DOI] [PubMed] [Google Scholar]
  6. Creuzet F., McDermott A., Gebhard R., van der Hoef K., Spijker-Assink M. B., Herzfeld J., Lugtenburg J., Levitt M. H., Griffin R. G. Determination of membrane protein structure by rotational resonance NMR: bacteriorhodopsin. Science. 1991 Feb 15;251(4995):783–786. doi: 10.1126/science.1990439. [DOI] [PubMed] [Google Scholar]
  7. Gasset M., Baldwin M. A., Lloyd D. H., Gabriel J. M., Holtzman D. M., Cohen F., Fletterick R., Prusiner S. B. Predicted alpha-helical regions of the prion protein when synthesized as peptides form amyloid. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10940–10944. doi: 10.1073/pnas.89.22.10940. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hsiao K. K., Groth D., Scott M., Yang S. L., Serban H., Rapp D., Foster D., Torchia M., Dearmond S. J., Prusiner S. B. Serial transmission in rodents of neurodegeneration from transgenic mice expressing mutant prion protein. Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):9126–9130. doi: 10.1073/pnas.91.19.9126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hsiao K. K., Scott M., Foster D., Groth D. F., DeArmond S. J., Prusiner S. B. Spontaneous neurodegeneration in transgenic mice with mutant prion protein. Science. 1990 Dec 14;250(4987):1587–1590. doi: 10.1126/science.1980379. [DOI] [PubMed] [Google Scholar]
  10. Huang Z., Prusiner S. B., Cohen F. E. Scrapie prions: a three-dimensional model of an infectious fragment. Fold Des. 1996;1(1):13–19. doi: 10.1016/S1359-0278(96)00007-7. [DOI] [PubMed] [Google Scholar]
  11. Kellings K., Meyer N., Mirenda C., Prusiner S. B., Riesner D. Further analysis of nucleic acids in purified scrapie prion preparations by improved return refocusing gel electrophoresis. J Gen Virol. 1992 Apr;73(Pt 4):1025–1029. doi: 10.1099/0022-1317-73-4-1025. [DOI] [PubMed] [Google Scholar]
  12. Lansbury P. T., Jr, Costa P. R., Griffiths J. M., Simon E. J., Auger M., Halverson K. J., Kocisko D. A., Hendsch Z. S., Ashburn T. T., Spencer R. G. Structural model for the beta-amyloid fibril based on interstrand alignment of an antiparallel-sheet comprising a C-terminal peptide. Nat Struct Biol. 1995 Nov;2(11):990–998. doi: 10.1038/nsb1195-990. [DOI] [PubMed] [Google Scholar]
  13. Meyer N., Rosenbaum V., Schmidt B., Gilles K., Mirenda C., Groth D., Prusiner S. B., Riesner D. Search for a putative scrapie genome in purified prion fractions reveals a paucity of nucleic acids. J Gen Virol. 1991 Jan;72(Pt 1):37–49. doi: 10.1099/0022-1317-72-1-37. [DOI] [PubMed] [Google Scholar]
  14. Nguyen J., Baldwin M. A., Cohen F. E., Prusiner S. B. Prion protein peptides induce alpha-helix to beta-sheet conformational transitions. Biochemistry. 1995 Apr 4;34(13):4186–4192. doi: 10.1021/bi00013a006. [DOI] [PubMed] [Google Scholar]
  15. Pan K. M., Baldwin M., Nguyen J., Gasset M., Serban A., Groth D., Mehlhorn I., Huang Z., Fletterick R. J., Cohen F. E. Conversion of alpha-helices into beta-sheets features in the formation of the scrapie prion proteins. Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):10962–10966. doi: 10.1073/pnas.90.23.10962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Prusiner S. B. Chemistry and biology of prions. Biochemistry. 1992 Dec 15;31(49):12277–12288. doi: 10.1021/bi00164a001. [DOI] [PubMed] [Google Scholar]
  17. Prusiner S. B., Groth D., Serban A., Koehler R., Foster D., Torchia M., Burton D., Yang S. L., DeArmond S. J. Ablation of the prion protein (PrP) gene in mice prevents scrapie and facilitates production of anti-PrP antibodies. Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10608–10612. doi: 10.1073/pnas.90.22.10608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Prusiner S. B., McKinley M. P., Bowman K. A., Bolton D. C., Bendheim P. E., Groth D. F., Glenner G. G. Scrapie prions aggregate to form amyloid-like birefringent rods. Cell. 1983 Dec;35(2 Pt 1):349–358. doi: 10.1016/0092-8674(83)90168-x. [DOI] [PubMed] [Google Scholar]
  19. Safar J., Roller P. P., Gajdusek D. C., Gibbs C. J., Jr Scrapie amyloid (prion) protein has the conformational characteristics of an aggregated molten globule folding intermediate. Biochemistry. 1994 Jul 12;33(27):8375–8383. doi: 10.1021/bi00193a027. [DOI] [PubMed] [Google Scholar]
  20. Smith S. O., Jonas R., Braiman M., Bormann B. J. Structure and orientation of the transmembrane domain of glycophorin A in lipid bilayers. Biochemistry. 1994 May 24;33(20):6334–6341. doi: 10.1021/bi00186a037. [DOI] [PubMed] [Google Scholar]
  21. Stahl N., Baldwin M. A., Teplow D. B., Hood L., Gibson B. W., Burlingame A. L., Prusiner S. B. Structural studies of the scrapie prion protein using mass spectrometry and amino acid sequencing. Biochemistry. 1993 Mar 2;32(8):1991–2002. doi: 10.1021/bi00059a016. [DOI] [PubMed] [Google Scholar]
  22. Stahl N., Borchelt D. R., Hsiao K., Prusiner S. B. Scrapie prion protein contains a phosphatidylinositol glycolipid. Cell. 1987 Oct 23;51(2):229–240. doi: 10.1016/0092-8674(87)90150-4. [DOI] [PubMed] [Google Scholar]
  23. Taraboulos A., Serban D., Prusiner S. B. Scrapie prion proteins accumulate in the cytoplasm of persistently infected cultured cells. J Cell Biol. 1990 Jun;110(6):2117–2132. doi: 10.1083/jcb.110.6.2117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Thompson L. K., McDermott A. E., Raap J., van der Wielen C. M., Lugtenburg J., Herzfeld J., Griffin R. G. Rotational resonance NMR study of the active site structure in bacteriorhodopsin: conformation of the Schiff base linkage. Biochemistry. 1992 Sep 1;31(34):7931–7938. doi: 10.1021/bi00149a026. [DOI] [PubMed] [Google Scholar]
  25. Wishart D. S., Sykes B. D. The 13C chemical-shift index: a simple method for the identification of protein secondary structure using 13C chemical-shift data. J Biomol NMR. 1994 Mar;4(2):171–180. doi: 10.1007/BF00175245. [DOI] [PubMed] [Google Scholar]
  26. Zhang H., Kaneko K., Nguyen J. T., Livshits T. L., Baldwin M. A., Cohen F. E., James T. L., Prusiner S. B. Conformational transitions in peptides containing two putative alpha-helices of the prion protein. J Mol Biol. 1995 Jul 21;250(4):514–526. doi: 10.1006/jmbi.1995.0395. [DOI] [PubMed] [Google Scholar]
  27. de Dios A. C., Pearson J. G., Oldfield E. Secondary and tertiary structural effects on protein NMR chemical shifts: an ab initio approach. Science. 1993 Jun 4;260(5113):1491–1496. doi: 10.1126/science.8502992. [DOI] [PubMed] [Google Scholar]

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