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首页> 美国卫生研究院文献>The Journal of Experimental Medicine >PHYSICAL CHEMICAL AND ULTRASTRUCTURAL STUDIES OF WATER-SOLUBLE HUMAN AMYLOID FIBRILS
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PHYSICAL CHEMICAL AND ULTRASTRUCTURAL STUDIES OF WATER-SOLUBLE HUMAN AMYLOID FIBRILS

机译：水溶性人淀粉样原纤维的物理化学和超结构研究

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Amyloid fibrils were isolated from the tissues of nine patients with amyloidosis in a state of high purity by homogenization of the tissue followed by extraction with distilled water. Physical, chemical, and ultrastructural studies suggest that amyloid fibrils from different individuals resemble each other, but are not identical. In tissue sections as well as by negative staining of isolated fibrils, morphologic variations were observed. Among the isolated fibrils at least three types were noted. The majority resembled those described previously. However, one subject had two types of fibrils which differed in size and appearance. Most of the preparations sedimented as a single component with a sedimentation coefficient of 45–50S or as a larger polymer. However, two of the preparations had sedimentation coefficients of 8–9S, and a third one had a major 95S component and a minor 9S fraction. While the preparations of amyloid were not sufficiently pure for amino acid analyses, peptide maps demonstrated differences among amyloid preparations from different subjects. The amyloid fibrils in their native state proved to be remarkably resistant to digestion by a number of proteolytic enzymes. Several chemical methods were tried to produce smaller subunits. Of these, the most successful one was the use of 0.1 M NaOH which yielded a smaller, soluble fraction with sedimentation coefficients ranging from 1.1 to 2.8S. Accompanying this degradation, there was little loss of peptides or carbohydrates. Based on the results of the chemical analyses, it is estimated that the subunit produced by sodium hydroxide had a molecular weight of approximately 35,000–40,000.
机译：通过将组织匀浆，然后用蒸馏水提取，以高纯度的状态从9名淀粉样变性患者的组织中分离出淀粉样原纤维。物理，化学和超微结构研究表明，来自不同个体的淀粉样蛋白原纤维彼此相似，但并不完全相同。在组织切片中以及通过对分离的原纤维进行负染色，观察到形态学变化。在分离的原纤维中，注意到至少三种类型。大多数类似于先前描述的内容。然而，一个受试者具有两种类型的原纤维，其大小和外观不同。大多数制剂以单个成分的形式沉淀，沉淀系数为45-50S或更大的聚合物。但是，其中两种制剂的沉降系数为8-9S，第三种制剂的主要成分为95S，次要成分为9S。虽然淀粉样蛋白制剂的纯度不足以进行氨基酸分析，但肽图显示了来自不同受试者的淀粉样蛋白制剂之间的差异。天然状态的淀粉样原纤维被证明对多种蛋白水解酶具有显着的抗消化性。尝试了几种化学方法来生产较小的亚基。其中，最成功的方法是使用0.1 M NaOH，生成的沉淀物系数从1.1到2.8S较小，可溶级分。伴随这种降解，肽或碳水化合物几乎没有损失。根据化学分析的结果，估计由氢氧化钠产生的亚基的分子量约为35,000–40,000。

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