在大肠杆菌中高效表达的完整成熟牛朊病毒蛋白：生化与结构研究

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

作者信息

Negro A, De Filippis V, Skaper S D, James P, Sorgato M C

机构信息

Dipartimento di Chimica Biologica, Centro CNR dello Studio delle Biomembrane, Università di Padova, Padua, Italy.

出版信息

FEBS Lett. 1997 Jul 28;412(2):359-64. doi: 10.1016/s0014-5793(97)00798-9.

DOI:10.1016/s0014-5793(97)00798-9

PMID:9256252

Abstract

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.

摘要

根据“仅蛋白质”假说，组成细胞的蛋白质PrP(C)的三维折叠结构转变为与疾病相关的异构体PrP(Sc)，是导致动物和人类神经退行性疾病的原因。在此，我们描述了在大肠杆菌中进行的高水平合成，以及以单体形式纯化的牛脑组氨酸标签全长成熟PrP（25-249），称为His-PrP。基于生化和光谱数据，His-PrP表现出PrP(C)异构体预期的特征。所报道的表达系统应能产生足够数量的牛PrP(C)，以允许进行三维结构测定。