Leliveld Sirik Rutger, Dame Remus Thei, Wuite Gijs J L, Stitz Lothar, Korth Carsten
Institute for Neuropathology, Heinrich Heine University of Düsseldorf, 40225 Düsseldorf, Germany.
J Biol Chem. 2006 Feb 10;281(6):3268-75. doi: 10.1074/jbc.M510606200. Epub 2005 Dec 13.
Insertion of additional octarepeats into the prion protein gene has been genetically linked to familial Creutzfeldt Jakob disease and hence to de novo generation of infectious prions. The pivotal event during prion formation is the conversion of the normal prion protein (PrPC) into the pathogenic conformer PrPSc, which subsequently induces further conversion in an autocatalytic manner. Apparently, an expanded octarepeat domain directs folding of PrP toward the PrPSc conformation and initiates a self-replicating conversion process. Here, based on three main observations, we have provided a model on how altered molecular interactions between wild-type and mutant PrP set the stage for familial Creutzfeldt Jakob disease with octarepeat insertions. First, we showed that wild-type octarepeat domains interact in a copper-dependent and reversible manner, a "copper switch." This interaction becomes irreversible upon domain expansion, possibly reflecting a loss of function. Second, expanded octarepeat domains of increasing length gradually form homogenous globular multimers of 11-21 nm in the absence of copper ions when expressed as soluble glutathione S-transferase fusion proteins. Third, octarepeat domain expansion causes a gain of function with at least 10 repeats selectively binding PrPSc in a denaturant-resistant complex in the absence of copper ions. Thus, the combination of both a loss and gain of function profoundly influences homomeric interaction behavior of PrP with an expanded octarepeat domain. A multimeric cluster of prion proteins carrying expanded octarepeat domains may therefore capture and incorporate spontaneously arising short-lived PrPSc-like conformers, thereby providing a matrix for their conversion.
在朊病毒蛋白基因中插入额外的八肽重复序列已在遗传学上与家族性克雅氏病相关联,从而与传染性朊病毒的新生有关。朊病毒形成过程中的关键事件是正常朊病毒蛋白(PrPC)转化为致病性构象体PrPSc,随后PrPSc以自催化方式诱导进一步转化。显然,扩展的八肽重复结构域将PrP的折叠导向PrPSc构象,并启动自我复制的转化过程。在此,基于三个主要观察结果,我们提供了一个模型,说明野生型和突变型PrP之间改变的分子相互作用如何为伴有八肽重复插入的家族性克雅氏病奠定基础。首先,我们表明野生型八肽重复结构域以铜依赖且可逆的方式相互作用,即“铜开关”。结构域扩展后这种相互作用变得不可逆,可能反映了功能丧失。其次,当作为可溶性谷胱甘肽S-转移酶融合蛋白表达时,长度增加的扩展八肽重复结构域在没有铜离子的情况下逐渐形成11-21纳米的均匀球状多聚体。第三,八肽重复结构域扩展导致功能获得,至少10个重复序列在没有铜离子的情况下能在抗变性剂复合物中选择性结合PrPSc。因此,功能丧失和获得的结合深刻影响了具有扩展八肽重复结构域的PrP的同聚相互作用行为。因此,携带扩展八肽重复结构域的朊病毒蛋白多聚体簇可能捕获并纳入自发产生的短寿命PrPSc样构象体,从而为它们的转化提供一个基质。