Oxidation of Helix-3 Methionines Precedes the Formation of PK Resistant PrPSc

PLoS Pathogens

Ruth Gabizon¹  Tamar Canello¹  Kati Frid¹  Jackob Moskovitz²  Assaf Friedler³  Ronen Gabizon³  María Gasset⁴  Silvia Lisa⁴ 

[1] Department of Neurology, The Agnes Ginges Center for Human Neurogenetics, Hadassah University Hospital, Jerusalem, Israel;Department of Pharmacology and Toxicology, School of Pharmacy, University of Kansas, Lawrence, United States of America;Institute of Chemistry, The Hebrew University of Jerusalem, Safra Campus, Jerusalem, Israel;Instituto Química-Física Rocasolano, Consejo Superior de Investigaciones Científicas, Madrid, Spain

关键词: Oxidation;    Proteases;    Animal prion diseases;    Antibodies;    Prion diseases;    Methionine;    Immunoblotting;    Scrapie;   

DOI  :  10.1371/journal.ppat.1000977

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

While elucidating the peculiar epitope of the α-PrP mAb IPC2, we found that PrPSc exhibits the sulfoxidation of residue M213 as a covalent signature. Subsequent computational analysis predicted that the presence of sulfoxide groups at both Met residues 206 and 213 destabilize the α-fold, suggesting oxidation may facilitate the conversion of PrPC into PrPSc. To further study the effect of oxidation on prion formation, we generated pAbs to linear PrP peptides encompassing the Helix-3 region, as opposed to the non-linear complexed epitope of IPC2. We now show that pAbs, whose epitopes comprise Met residues, readily detected PrPC, but could not recognize most PrPSc bands unless they were vigorously reduced. Next, we showed that the α-Met pAbs did not recognize newly formed PrPSc, as is the case for the PK resistant PrP present in lines of prion infected cells. In addition, these reagents did not detect intermediate forms such as PK sensitive and partially aggregated PrPs present in infected brains. Finally, we show that PrP molecules harboring the pathogenic mutation E200K, which is linked to the most common form of familial CJD, may be spontaneously oxidized. We conclude that the oxidation of methionine residues in Helix-3 represents an early and important event in the conversion of PrPC to PrPSc. We believe that further investigation into the mechanism and role of PrP oxidation will be central in finally elucidating the mechanism by which a normal cell protein converts into a pathogenic entity that causes fatal brain degeneration.

【 授权许可】

CC BY