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NMR structure of the human prion protein with the pathological Q212P mutation reveals unique structural features.

Ilc G, Giachin G, Jaremko M, Jaremko Ł, Benetti F, Plavec J, Zhukov I, Legname G - PLoS ONE (2010)

Bottom Line: The substitution of a glutamine by a proline at the position 212 introduces novel structural differences in comparison to the known wild-type PrP structures.This structure might provide new insights into the early events of conformational transition of PrP(C) into PrP(Sc).Indeed, the spontaneous formation of prions in familial cases might be due to the disruptions of the hydrophobic core consisting of beta(2)-alpha(2) loop and alpha(3) helix.

View Article: PubMed Central - PubMed

Affiliation: Slovenian NMR Centre, National Institute of Chemistry, Ljubljana, Slovenia.

ABSTRACT
Prion diseases are fatal neurodegenerative disorders caused by an aberrant accumulation of the misfolded cellular prion protein (PrP(C)) conformer, denoted as infectious scrapie isoform or PrP(Sc). In inherited human prion diseases, mutations in the open reading frame of the PrP gene (PRNP) are hypothesized to favor spontaneous generation of PrP(Sc) in specific brain regions leading to neuronal cell degeneration and death. Here, we describe the NMR solution structure of the truncated recombinant human PrP from residue 90 to 231 carrying the Q212P mutation, which is believed to cause Gerstmann-Sträussler-Scheinker (GSS) syndrome, a familial prion disease. The secondary structure of the Q212P mutant consists of a flexible disordered tail (residues 90-124) and a globular domain (residues 125-231). The substitution of a glutamine by a proline at the position 212 introduces novel structural differences in comparison to the known wild-type PrP structures. The most remarkable differences involve the C-terminal end of the protein and the beta(2)-alpha(2) loop region. This structure might provide new insights into the early events of conformational transition of PrP(C) into PrP(Sc). Indeed, the spontaneous formation of prions in familial cases might be due to the disruptions of the hydrophobic core consisting of beta(2)-alpha(2) loop and alpha(3) helix.

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NMR spectra and assignment of HuPrP(90–231, M129, Q212P).(A) 1H-15N HSQC spectrum with one letter amino acid code. (B) Backbone assignment for the part that shows unique structural features. Strips for residues 221-230 from 3D CA(CO)NH and HNCA experiments with cross-peaks in black from 13Cα(i-1) nuclei in CA(CO)NH experiment and in red corresponding to 13Cα(i-1) and 13Cα(i) nuclei revealed by HNCA experiment. Blue line indicates sequential walk.
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pone-0011715-g001: NMR spectra and assignment of HuPrP(90–231, M129, Q212P).(A) 1H-15N HSQC spectrum with one letter amino acid code. (B) Backbone assignment for the part that shows unique structural features. Strips for residues 221-230 from 3D CA(CO)NH and HNCA experiments with cross-peaks in black from 13Cα(i-1) nuclei in CA(CO)NH experiment and in red corresponding to 13Cα(i-1) and 13Cα(i) nuclei revealed by HNCA experiment. Blue line indicates sequential walk.

Mentions: 1H-15N HSQC spectrum of 13C, 15N double labeled mutant demonstrated good dispersion of amide signals (Figure 1A). Cross-peaks for all residues could be identified with the exception of Arg164, Tyr169, Asn171, Phe175 and Gln217, which were not observed due to line broadening caused by exchange processes (see below). The sequence-specific assignment was achieved with the use of standard triple resonance HNCA, HN(CO)CA, HNCACB and CBCA(CO)NH NMR experiments. Strips from 3D CA(CO)NH and HNCA experiments in Figure 1B illustrate sequential walk in the region from Glu221 to Ser230. The assignment was additionally confirmed by analysis of sequential and medium-range NOEs in 3D 15N-edited NOESY-HSQC experiment. 1H and 13C resonances of side chains were assigned by analyses of 3D (H)CCH-TOCSY and 13C-edited NOESY-HSQC spectra. Final level of completeness of 1H, 13C and 15N resonance assignment was very high (95.1%). Chemical shifts were deposited in BioMagnetic Resonance data Bank (BMRB, accession code 16743).


NMR structure of the human prion protein with the pathological Q212P mutation reveals unique structural features.

Ilc G, Giachin G, Jaremko M, Jaremko Ł, Benetti F, Plavec J, Zhukov I, Legname G - PLoS ONE (2010)

NMR spectra and assignment of HuPrP(90–231, M129, Q212P).(A) 1H-15N HSQC spectrum with one letter amino acid code. (B) Backbone assignment for the part that shows unique structural features. Strips for residues 221-230 from 3D CA(CO)NH and HNCA experiments with cross-peaks in black from 13Cα(i-1) nuclei in CA(CO)NH experiment and in red corresponding to 13Cα(i-1) and 13Cα(i) nuclei revealed by HNCA experiment. Blue line indicates sequential walk.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2908606&req=5

pone-0011715-g001: NMR spectra and assignment of HuPrP(90–231, M129, Q212P).(A) 1H-15N HSQC spectrum with one letter amino acid code. (B) Backbone assignment for the part that shows unique structural features. Strips for residues 221-230 from 3D CA(CO)NH and HNCA experiments with cross-peaks in black from 13Cα(i-1) nuclei in CA(CO)NH experiment and in red corresponding to 13Cα(i-1) and 13Cα(i) nuclei revealed by HNCA experiment. Blue line indicates sequential walk.
Mentions: 1H-15N HSQC spectrum of 13C, 15N double labeled mutant demonstrated good dispersion of amide signals (Figure 1A). Cross-peaks for all residues could be identified with the exception of Arg164, Tyr169, Asn171, Phe175 and Gln217, which were not observed due to line broadening caused by exchange processes (see below). The sequence-specific assignment was achieved with the use of standard triple resonance HNCA, HN(CO)CA, HNCACB and CBCA(CO)NH NMR experiments. Strips from 3D CA(CO)NH and HNCA experiments in Figure 1B illustrate sequential walk in the region from Glu221 to Ser230. The assignment was additionally confirmed by analysis of sequential and medium-range NOEs in 3D 15N-edited NOESY-HSQC experiment. 1H and 13C resonances of side chains were assigned by analyses of 3D (H)CCH-TOCSY and 13C-edited NOESY-HSQC spectra. Final level of completeness of 1H, 13C and 15N resonance assignment was very high (95.1%). Chemical shifts were deposited in BioMagnetic Resonance data Bank (BMRB, accession code 16743).

Bottom Line: The substitution of a glutamine by a proline at the position 212 introduces novel structural differences in comparison to the known wild-type PrP structures.This structure might provide new insights into the early events of conformational transition of PrP(C) into PrP(Sc).Indeed, the spontaneous formation of prions in familial cases might be due to the disruptions of the hydrophobic core consisting of beta(2)-alpha(2) loop and alpha(3) helix.

View Article: PubMed Central - PubMed

Affiliation: Slovenian NMR Centre, National Institute of Chemistry, Ljubljana, Slovenia.

ABSTRACT
Prion diseases are fatal neurodegenerative disorders caused by an aberrant accumulation of the misfolded cellular prion protein (PrP(C)) conformer, denoted as infectious scrapie isoform or PrP(Sc). In inherited human prion diseases, mutations in the open reading frame of the PrP gene (PRNP) are hypothesized to favor spontaneous generation of PrP(Sc) in specific brain regions leading to neuronal cell degeneration and death. Here, we describe the NMR solution structure of the truncated recombinant human PrP from residue 90 to 231 carrying the Q212P mutation, which is believed to cause Gerstmann-Sträussler-Scheinker (GSS) syndrome, a familial prion disease. The secondary structure of the Q212P mutant consists of a flexible disordered tail (residues 90-124) and a globular domain (residues 125-231). The substitution of a glutamine by a proline at the position 212 introduces novel structural differences in comparison to the known wild-type PrP structures. The most remarkable differences involve the C-terminal end of the protein and the beta(2)-alpha(2) loop region. This structure might provide new insights into the early events of conformational transition of PrP(C) into PrP(Sc). Indeed, the spontaneous formation of prions in familial cases might be due to the disruptions of the hydrophobic core consisting of beta(2)-alpha(2) loop and alpha(3) helix.

Show MeSH
Related in: MedlinePlus