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The structural intolerance of the PrP α-fold for polar substitution of the helix-3 methionines

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Abstract

The conversion of the cellular prion protein (PrPC) into its disease-associated form (PrPSc) involves a major conformational change and the accumulation of sulfoxidized methionines. Computational and synthetic approaches have shown that this change in the polarity of M206 and M213 impacts the C-terminal domain native α-fold allowing the flexibility required for the structural conversion. To test the effect in the full-length molecule with site-specificity, we have generated M-to-S mutations. Molecular dynamics simulations show that the replacement indeed perturbs the native state. When this mutation is placed at the conserved methionines of HaPrP(23–231), only substitutions at the Helix-3 impair the α-fold, stabilizing a non-native state with perturbed secondary structure, loss of native tertiary contacts, increased surface hydrophobicity, reduced thermal stability and an enhanced tendency to aggregate into protofibrillar polymers. Our work supports that M206 and M213 function as α-fold gatekeepers and suggests that their redox state regulate misfolding routes.

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Abbreviations

PrPC :

Cellular prion protein

PrPSc :

Disease-related form of PrP

HuPrP(125–229):

Polypeptide chain representing the globular domain of the human PrP

HaPrP(23–231):

Polypeptide chain representing the mature chain of the hamster PrP

MD:

Molecular dynamics

RWISP:

Root weighted square inner product

CP:

Communication propensity

CD:

Circular dichroism

DLS:

Dynamic light scattering

Rh :

Hydrodynamic radius

ThT:

Thioflavin T

AFM:

Atomic force microscopy

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Acknowledgments

This work was supported by grants SAF2006-00418 (MG) and BFU2009-07971 (MG) from the Ministerio de Ciencia e Innovación, FOOD-CT-2004-506579 (MG, RG) from the EC and PI101209 (MG) from the Fundación Cien. SL is supported by a FPI-PhD fellowship from the Ministerio de Ciencia e Innovación. We gratefully acknowledge the advice of Dr. Angel Cuesta in relation to the AFM experiments and the technical support of Lara Reviejo and Rosa Sánchez.

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Authors and Affiliations

  1. Insto Química-Física “Rocasolano”, CSIC, Serrano 119, 28006, Madrid, Spain

    Silvia Lisa, Gema Cabello & María Gasset

  2. Istituto di Chimica del Riconoscimento Molecolare, Consiglio Nazionale delle Ricerche, Via Mario Bianco 9, 20131, Milan, Italy

    Massimiliano Meli & Giorgio Colombo

  3. Department of Neurology, The Agnes Ginges Center for Human Neurogenetics, Hadassah University Hospital, 91120, Jerusalem, Israel

    Ruth Gabizon

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  1. Silvia Lisa
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  2. Massimiliano Meli
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  3. Gema Cabello
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  4. Ruth Gabizon
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  5. Giorgio Colombo
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  6. María Gasset
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Correspondence to María Gasset.

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S. Lisa and M. Meli contributed equally to this work.

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Lisa, S., Meli, M., Cabello, G. et al. The structural intolerance of the PrP α-fold for polar substitution of the helix-3 methionines. Cell. Mol. Life Sci. 67, 2825–2838 (2010). https://doi.org/10.1007/s00018-010-0363-1

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