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Measurement of amyloid fibril mass-per-length by tilted-beam transmission electron microscopy

机译:倾斜电子束透射电子显微镜测量淀粉样蛋白原纤维质量

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摘要

We demonstrate that accurate values of mass-per-length (MPL), which serve as strong constraints on molecular structure, can be determined for amyloid fibrils by quantification of intensities in dark-field electron microscope images obtained in the tilted-beam mode of a transmission electron microscope. MPL values for fibrils formed by residues 218-289 of the HET-s fungal prion protein, for 2-fold- and 3-fold-symmetric fibrils formed by the 40-residue β-amyloid peptide, and for fibrils formed by the yeast prion protein Sup35NM are in good agreement with previous results from scanning transmission electron microscopy. Results for fibrils formed by the yeast prion protein Rnq1, for which the MPL value has not been previously reported, support an in-register parallel β-sheet structure, with one Rnq1 molecule per 0.47-nm β-sheet repeat spacing. Since tilted-beam dark-field images can be obtained on many transmission electron microscopes, this work should facilitate MPL determination by a large number of research groups engaged in studies of amyloid fibrils and similar supramo-lecular assemblies.
机译:我们证明,可以通过量化在倾斜光束模式下获得的暗场电子显微镜图像中的强度来确定淀粉样蛋白原纤维的精确值,即对分子结构有严格限制的质量平均长度(MPL)。透射电子显微镜。由HET-s真菌pr病毒蛋白残基218-289形成的原纤维,由40个残基的β-淀粉样肽形成的2倍和3倍对称原纤维以及由酵母病毒形成的原纤维的MPL值Sup35NM蛋白与扫描透射电子显微镜的先前结果非常吻合。由酵母pr病毒蛋白Rnq1形成的原纤维(其MPL值先前尚未报道)的结果支持寄存器内平行β-折叠结构,每0.47 nmβ-折叠重复间隔一个Rnq1分子。由于可以在许多透射电子显微镜上获得倾斜光束暗场图像,因此这项工作应有助于从事淀粉样蛋白原纤维和类似超分子-分子组装研究的许多研究小组对MPL的测定。

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  • 作者单位

    Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892;

    Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892;

    Laboratory of Biochemistry and Genetics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892;

    Laboratory of Biochemistry and Genetics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892;

    Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    alzheimer's disease; molecular structure; prions; solid state NMR;

    机译:阿尔茨海默氏病;分子结构ions病毒;固态NMR;

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