The complete mature bovine prion protein highly expressed in Escherichia coli: biochemical and structural studies

FEBS Letters

Skaper, Stephen D¹  De Filippis, Vincenzo³  Sorgato, M.Catia²  Negro, Alessandro² 

[1] Dipartimento di Farmacologia, Università di Padova, Padua, Italy;Dipartimento di Chimica Biologica, Centro CNR dello Studio delle Biomembrane, Università di Padova, Viale G. Colombo 3, 35121 Padua, Italy;CRIBI Biotechnology Center, Università di Padova, Padua, Italy

关键词: Spongiform encephalopathy;    Scrapie;    Prion;    Recombinant bovine prion protein;    PrP;    prion protein;    PrPC;    cellular form of the prion protein;    PrPSc;    pathogenic form of the prion protein;    HPLC;    high-performance liquid chromatography;    CD;    circular dichroism;    SDS-PAGE;    sodium dodecylsulfate-polyacrylamide gel electrophoresis;    TFA;    trifluoroacetic acid;    PCR;    polymerase chain reaction;    PVDF;    polyvinylidene difluoride;   

DOI  :  10.1016/S0014-5793(97)00798-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

According to the `protein only' hypothesis, modification of the 3-dimensional fold of the constituent cellular protein, PrP^C, into the disease-associated isoform, PrP^Sc, is the cause of neurodegenerative diseases in animals and humans. Here we describe the high-level synthesis in Escherichia coli, and purification in the monomeric form, of a histidine-tagged full-length mature PrP (25–249) of bovine brain, termed His-PrP. Based on biochemical and spectroscopic data, His-PrP displays characteristics expected for the PrP^C isoform. The reported expression system should allow the production of quantities of bovine PrP^C sufficient to permit 3-dimensional structure determinations.

【 授权许可】

Unknown