Using small molecule reagents to selectively modify epitopes based on their conformation.
Using small molecule reagents to selectively modify epitopes based on their conformation.
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作者：
Silva CJ
PrPSc是一种感染性蛋白。唯一实验证实的差异之间的PrPSc和正常的细胞异构体（PrPC）是构象。这项工作描述了一种确定存在于PrPSc异构体中的表面暴露或隔离氨基酸的方法。合成了n -羟基琥珀酰亚胺乙酸酯和4-三甲氨基丁酸酯，并与me7感染小鼠、未感染小鼠、263k感染仓鼠和未感染仓鼠的脑提取物进行了洗涤剂溶解反应。用抗体3F4、6D11、7D9、AG4、AH6、GE8或MAB5424检测这些反应混合物。3F4, 6D11， AH6和GE8抗体识别在PrPSc异构体中加密但在PrPC异构体中暴露的表位。这些试剂允许检测朊病毒感染的脑提取物，而不需要蛋白酶K消化。此外，它们可以与适当的抗体一起使用，以确定PrPSc的哪些氨基酸暴露在表面，哪些被加密，从而提供有用的结构信息。在不使用蛋白酶K的情况下，这种方法被用来区分263K和困倦的仓鼠适应痒病菌株。
PrPSc is an infectious protein. The only experimentally verified difference between PrPSc and its normal cellular isoform (PrPC) is conformational. This work describes an approach to determining the presence of surface exposed or sequestered amino acids present in the PrPSc isoform. The N-hydroxysuccinimide esters of acetic acid and 4-trimethylammoniumbutyric acid were synthesized and reacted with detergent-solubilized brain extracts from Me7-infected mice, uninfected mice, 263K-infected hamsters or uninfected hamsters. These reaction mixtures were analyzed by western blots probed with the antibodies 3F4, 6D11, 7D9, AG4, AH6, GE8 or MAB5424. The 3F4, 6D11, AH6, and GE8 antibodies recognize an epitope that is encrypted in the PrPSc isoform, but exposed in the PrPC isoform. These reagents permit the detection of prion infected brain extracts without the need for proteinase K digestion. In addition they can be used, with an appropriate antibody, to determine which amino acids of PrPSc are exposed on the surface and which are encrypted, thus providing useful structural information. This approach was used to distinguish between the 263K and drowsy strains of hamster-adapted scrapie without the use of proteinase K.
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影响因子：
8
作者：
Peretz, D;Scott, MR;Prusiner, SB
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Prusiner, SB
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作者：
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通讯作者：
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