Caughey B, Raymond G J, Bessen R A
Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, NIAID, National Institutes of Health, Hamilton, Montana 59840, USA.
J Biol Chem. 1998 Nov 27;273(48):32230-5. doi: 10.1074/jbc.273.48.32230.
Strain diversity in the transmissible spongiform encephalopathies (TSEs) has been proposed to be determined by variations in the conformation of the abnormal, protease-resistant form of prion protein (PrP-res). We have investigated whether infection of hamsters with three TSE strains resulted in the formation of PrP-res with different conformations using limited proteinase K (PK) digestion and infrared spectroscopy. PrP-res isolated from the brains of hamsters infected with the hyper (HY), drowsy (DY), and 263K TSE strains yielded similar SDS-polyacrylamide gel electrophoresis profiles prior to PK treatment. However, after limited digestion with PK, the PrP-res from the DY strain exhibited a fragmentation pattern that was distinct from that of the other two strains. Infrared spectra of HY and 263K PrP-res each had major absorption bands in the amide I region at 1626 and 1636 cm-1 both prior to and after digestion with PK. These bands were not evident in the DY PrP-res spectra, which had a unique band at 1629-1630 cm-1 and stronger band intensity at both 1616 and 1694-1695 cm-1. Because absorbances from 1616 to 1636 cm-1 of protein infrared spectra are attributed primarily to beta-sheet structures, these findings indicate that the conformations of HY and 263K PrP-res differ from DY PrP-res at least in structural regions with beta-sheet secondary structure. These results support the hypothesis that strain-specific PrP-res conformers can self-propagate by converting the normal prion protein to the abnormal conformers that induce phenotypically distinct TSE diseases.
有人提出,传染性海绵状脑病(TSEs)中的毒株多样性是由异常的、抗蛋白酶的朊病毒蛋白形式(PrP-res)构象变化所决定的。我们使用有限的蛋白酶K(PK)消化和红外光谱法,研究了用三种TSE毒株感染仓鼠是否会导致形成具有不同构象的PrP-res。在PK处理之前,从感染了高毒株(HY)、嗜睡毒株(DY)和263K TSE毒株的仓鼠大脑中分离出的PrP-res产生了相似的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳图谱。然而,在用PK进行有限消化后,DY毒株的PrP-res呈现出一种与其他两种毒株不同的片段化模式。HY和263K PrP-res的红外光谱在PK消化前后,酰胺I区域均在1626和1636 cm-1处有主要吸收带。这些吸收带在DY PrP-res光谱中不明显,DY PrP-res光谱在1629 - 1630 cm-1处有一个独特的吸收带,在1616和1694 - 1695 cm-1处有更强的吸收带强度。由于蛋白质红外光谱中1616至1636 cm-1的吸光度主要归因于β-折叠结构,这些发现表明,HY和263K PrP-res的构象至少在具有β-折叠二级结构的结构区域与DY PrP-res不同。这些结果支持了这样一种假说,即毒株特异性的PrP-res构象异构体可以通过将正常的朊病毒蛋白转化为诱导表型不同的TSE疾病的异常构象异构体来自我增殖。