TY - JOUR
T1 - Molecular properties of complexes formed between the prion protein and synthetic peptides
AU - Kaneko, Kiyotoshi
AU - Wille, Holger
AU - Mehlhorn, Ingrid
AU - Zhang, Hong
AU - Ball, Haydn
AU - Cohen, Fred E.
AU - Baldwin, Michael A.
AU - Prusiner, Stanley B.
N1 - Funding Information:
Supported by grants from the National Institutes of Health (NS14069, AG08967, AG02132, NS22786 and AG10770) and the American Health Assistance Foundation, as well as by gifts from the Sherman Fairchild Foundation and Bernard Osher Foundation. K. K. was supported in part by the Uehara Memorial Foundation and the Japanese Ministry of Education. H. W. was supported in part by a Feodor-Lynen-Fellowship of the Alexander von Humboldt-Stiftung. I. M. was supported by a senior fellowship from the Medical Research Council of Canada and by the French Foundation. ESI mass spectrometry was carried out in the UCSF Mass Spectrometry Facility (A. L. Burlingame, Director) supported by NIH NCRR BIPT RR01614.
PY - 1997/7/25
Y1 - 1997/7/25
N2 - Complexes of the Syrian hamster cellular prion protein (PrP(C)) and synthetic Syrian hamster PrP peptides were found to mimic many of the characteristics of Me scrapie PrP isoform (PrP(SC)). Either PrP(C) expressed in chinese hamster ovary (CHO) cells or a C-terminal fragment of 142 residues of recombinant PrP protein (rPrP) produced in Escherichia coli was mixed with an excess of a synthetic 56 amino acid peptide, denoted PrP(90-145). Complex formation required PrP(C) or rPrP to be destabilized by guanidine hydrochloride (GdnHCl) or urea and PrP(90-145) to be in a coil conformation; it was enhanced by an acidic environment, salt and detergent. If PrP(90-145) was in a β-sheet conformation, then no complexes were formed. While complex formation was rapid, acquisition' of protease resistance was a slow process. Amorphous aggregates with a PrP(C)/PrP(90-145) ratio of 1:1 were formed in phosphate buffer, whereas fibrils with a diameter of ~ 10 nm and a PrP(C)/PrP(90-145) ratio of 1:5 were formed in Tris buffer. The complexes were stable only in the presence of excess peptide in either the coil or β-sheet conformation; they dissociated rapidly after centrifugation and resuspension in buffer without peptide. Neither a peptide having a similar hydrophobicity profile/ charge distribution to PrP(90-145) nor a scrambled version, denoted hPrP(90-145) and sPrP(90-145), respectively, were able to induce complex formation. Although hPrP(90-145) could stabilize the PrP(C)/PrP(90-145) complexes, sPrP(90-145) could not. Studies of PrP(C)/peptide complexes may provide insights into how PrP(C) interacts with PrP(Sc) during the formation of a nascent PrP(Sc) molecule and into the process by which PrP(C) is converted into PrP(Sc).
AB - Complexes of the Syrian hamster cellular prion protein (PrP(C)) and synthetic Syrian hamster PrP peptides were found to mimic many of the characteristics of Me scrapie PrP isoform (PrP(SC)). Either PrP(C) expressed in chinese hamster ovary (CHO) cells or a C-terminal fragment of 142 residues of recombinant PrP protein (rPrP) produced in Escherichia coli was mixed with an excess of a synthetic 56 amino acid peptide, denoted PrP(90-145). Complex formation required PrP(C) or rPrP to be destabilized by guanidine hydrochloride (GdnHCl) or urea and PrP(90-145) to be in a coil conformation; it was enhanced by an acidic environment, salt and detergent. If PrP(90-145) was in a β-sheet conformation, then no complexes were formed. While complex formation was rapid, acquisition' of protease resistance was a slow process. Amorphous aggregates with a PrP(C)/PrP(90-145) ratio of 1:1 were formed in phosphate buffer, whereas fibrils with a diameter of ~ 10 nm and a PrP(C)/PrP(90-145) ratio of 1:5 were formed in Tris buffer. The complexes were stable only in the presence of excess peptide in either the coil or β-sheet conformation; they dissociated rapidly after centrifugation and resuspension in buffer without peptide. Neither a peptide having a similar hydrophobicity profile/ charge distribution to PrP(90-145) nor a scrambled version, denoted hPrP(90-145) and sPrP(90-145), respectively, were able to induce complex formation. Although hPrP(90-145) could stabilize the PrP(C)/PrP(90-145) complexes, sPrP(90-145) could not. Studies of PrP(C)/peptide complexes may provide insights into how PrP(C) interacts with PrP(Sc) during the formation of a nascent PrP(Sc) molecule and into the process by which PrP(C) is converted into PrP(Sc).
KW - Folding intermediate
KW - Insolubility
KW - Prion protein synthetic peptide
KW - Scrapie
KW - β-pleated sheet
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U2 - 10.1006/jmbi.1997.1135
DO - 10.1006/jmbi.1997.1135
M3 - Article
C2 - 9245588
AN - SCOPUS:0031586212
SN - 0022-2836
VL - 270
SP - 574
EP - 586
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 4
ER -