Protease-sensitive synthetic prions.
Protease-sensitive synthetic prions.
复制标题
DOI：
10.1371/journal.ppat.1000736
复制
发表时间：
2010-01-22
期刊：
影响因子：
6.7
通讯作者：
Prusiner SB
中科院分区：
文献类型：
--
作者：
Colby DW;Wain R;Baskakov IV;Legname G;Palmer CG;Nguyen HO;Lemus A;Cohen FE;DeArmond SJ;Prusiner SB
当细胞朊病毒蛋白（PrPC）经历自我传播的构象变化时，朊病毒出现;产生的感染性构象被命名为PrPSc。通常，PrPSc对蛋白酶具有耐药性，但已在人类和其他动物中分离出对蛋白酶敏感的朊病毒。我们在这里报告，蛋白酶敏感，合成朊病毒在体外聚合过程中产生的重组（rec）PrP成淀粉样纤维。在22个独立的实验中，recPrP淀粉样蛋白制剂，而不是recPrP单体或寡聚体，传播疾病的转基因小鼠（n = 164），表示Tg 9949小鼠，过表达N-末端截短的PrP。 Tg 9949对照小鼠（n = 174）没有自发产生朊病毒，尽管它们倾向于迟发性自发神经功能障碍。 当从感染的Tg 9949小鼠中分离的合成朊病毒在同一系小鼠中连续传播时，它们表现出sPrPSc并引起神经变性。有趣的是，这些蛋白酶敏感的朊病毒并没有缩短Tg 9949小鼠的寿命，尽管引起广泛的神经退行性变。我们将三种合成朊病毒分离株接种到过表达全长PrP的Tg 4053小鼠中; Tg 4053小鼠不易发生自发性神经功能障碍。合成的朊病毒分离物在600-750天内在表现出sPrPSc的Tg 4053小鼠中引起疾病。这些新的合成朊病毒表明，野生型朊蛋白的构象变化可以产生小鼠朊病毒组成的sPrPSc。朊病毒是一种感染性蛋白质，可引起人类和其他哺乳动物的遗传性、散发性和传播性疾病。当朊病毒蛋白（PrP）的正常形式采用自我永存构象时，这些感染性蛋白质就会出现。这种致病的PrP形式通常与正常PrP的区别在于其对蛋白酶消化的抵抗力，尽管大量证据表明蛋白酶敏感的朊病毒天然存在于人类和绵羊中。在这里，我们描述了新的蛋白酶敏感的合成朊病毒的一代。在大肠杆菌中产生野生型小鼠序列的重组PrP后，我们将蛋白质聚合成淀粉样纤维构象。接种这些淀粉样纤维的小鼠出现了朊病毒病的广泛神经变性特征，但没有产生蛋白酶抗性PrP。来自患病动物的朊病毒被传递给健康动物，健康动物同样发生神经变性，但不产生抗蛋白酶的朊病毒。这些新的合成朊病毒表明，截短的野生型PrP可以经历构象变化，变得具有感染性，但蛋白质仍然对蛋白酶敏感。
Prions arise when the cellular prion protein (PrPC) undergoes a self-propagating conformational change; the resulting infectious conformer is designated PrPSc. Frequently, PrPSc is protease-resistant but protease-sensitive (s) prions have been isolated in humans and other animals. We report here that protease-sensitive, synthetic prions were generated in vitro during polymerization of recombinant (rec) PrP into amyloid fibers. In 22 independent experiments, recPrP amyloid preparations, but not recPrP monomers or oligomers, transmitted disease to transgenic mice (n = 164), denoted Tg9949 mice, that overexpress N-terminally truncated PrP. Tg9949 control mice (n = 174) did not spontaneously generate prions although they were prone to late-onset spontaneous neurological dysfunction. When synthetic prion isolates from infected Tg9949 mice were serially transmitted in the same line of mice, they exhibited sPrPSc and caused neurodegeneration. Interestingly, these protease-sensitive prions did not shorten the life span of Tg9949 mice despite causing extensive neurodegeneration. We inoculated three synthetic prion isolates into Tg4053 mice that overexpress full-length PrP; Tg4053 mice are not prone to developing spontaneous neurological dysfunction. The synthetic prion isolates caused disease in 600–750 days in Tg4053 mice, which exhibited sPrPSc. These novel synthetic prions demonstrate that conformational changes in wild-type PrP can produce mouse prions composed exclusively of sPrPSc. Prions are infectious proteins that cause heritable, sporadic, and transmissible diseases in humans and other mammals. These infectious proteins arise when the normal form of the prion protein (PrP) adopts a self-perpetuating conformation. This disease-causing PrP form is frequently distinguished from normal PrP by its resistance to digestion by proteases although considerable evidence shows that protease-sensitive prions occur naturally in humans and sheep. Here we describe the generation of novel protease-sensitive synthetic prions. After producing recombinant PrP of the wild-type mouse sequence in Escherichia coli, we polymerized the protein into an amyloid fiber conformation. Mice inoculated with these amyloid fibers developed extensive neurodegeneration characteristic of prion disease, but did not generate protease-resistant PrP. Prions from sick animals were transmitted to healthy animals, which likewise developed neurodegeneration but not protease-resistant prions. These novel synthetic prions demonstrate that truncated wild-type PrP can undergo a conformational change that becomes infectious yet the protein remains protease sensitive.
登录
查看更多内容
影响因子：
5.6
作者：
Kaneko, K;Ball, HL;Cohen, FE
通讯作者：
Cohen, FE
DOI：
10.1073/pnas.0608970103
发表时间：
2006-12-12
影响因子：
11.1
作者：
Legname, Giuseppe;Nguyen, Hoang-Oanh B.;Prusiner, Stanley B.
通讯作者：
Prusiner, Stanley B.
DOI：
10.1073/pnas.83.8.2310
发表时间：
1986-04-01
影响因子：
11.1
作者：
MEYER, RK;MCKINLEY, MP;PRUSINER, SB
通讯作者：
PRUSINER, SB
影响因子：
82.9
作者：
Muramoto, T;DeArmond, SJ;Prusiner, SB
通讯作者：
Prusiner, SB
影响因子：
16.6
作者：
Caughey B;Baron GS;Chesebro B;Jeffrey M
通讯作者：
Jeffrey M