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首页> 美国卫生研究院文献>Prion >Applying the tools of chemistry (mass spectrometry and covalent modification by small molecule reagents) to the detection of prions and the study of their structure
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Applying the tools of chemistry (mass spectrometry and covalent modification by small molecule reagents) to the detection of prions and the study of their structure

机译：将化学工具（质谱分析和小分子试剂的共价修饰）应用于to病毒的检测及其结构的研究

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摘要

Prions are molecular pathogens, able to convert a normal cellular prion protein (PrP ) into a prion (PrP ). The information necessary for this conversion is contained in the conformation of PrP . Mass spectrometry (MS) and small-molecule covalent reactions have been used to study prions. Mass spectrometry has been used to detect and quantitate prions in the attomole range (10 mole). MS-based analysis showed that both possess identical amino acid sequences, one disulfide bond, a GPI anchor, asparagine-linked sugar antennae, and unoxidized methionines. Mass spectrometry has been used to define elements of the secondary and tertiary structure of wild-type PrP and GPI-anchorless PrP . It has also been used to study the quaternary structure of the PrP multimer. Small molecule reagents react differently with the same lysine in the PrP conformation than in the PrP conformation. Such differences can be detected by Western blot using mAbs with lysine-containing epitopes, such as 3F4 and 6D11. This permits the detection of PrP without the need for proteinase K pretreatment and can be used to distinguish among prion strains. These results illustrate how two important chemical tools, mass spectrometry and covalent modification by small molecules, are being applied to the detection and structural study of prions. Furthermore these tools are or can be applied to the study of the other protein misfolding diseases such as Alzheimer Disease, Parkinson Disease, or ALS.
机译：ions病毒是分子病原体，能够将正常的细胞病毒蛋白（PrP）转化为a病毒（PrP）。转换所需的信息包含在PrP的构象中。质谱（MS）和小分子共价反应已用于研究病毒。质谱法已被用于检测和定量在attomole范围（10摩尔）内的病毒。基于质谱的分析表明，两者都具有相同的氨基酸序列，一个二硫键，一个GPI锚，天冬酰胺连接的糖触角和未氧化的蛋氨酸。质谱已用于定义野生型PrP和GPI-无锚PrP的二级和三级结构的元素。它还已用于研究PrP多聚体的四级结构。小分子试剂与同一赖氨酸在PrP构象中的反应不同于在PrP构象中。可以使用具有赖氨酸表位（例如3F4和6D11）的mAb通过Western blot检测这种差异。这允许检测PrP，而无需进行蛋白酶K预处理，并可用于区分病毒菌株。这些结果说明了如何将两种重要的化学工具质谱和小分子共价修饰应用于modification病毒的检测和结构研究。此外，这些工具已经或可以用于研究其他蛋白质错误折叠疾病，例如阿尔茨海默氏病，帕金森氏病或ALS。

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