Bates I R, Matharu P, Ishiyama N, Rochon D, Wood D D, Polverini E, Moscarello M A, Viner N J, Harauz G
Department of Molecular Biology and Genetics, University of Guelph, Guelph, Ontario, N1G 2W1, Canada.
Protein Expr Purif. 2000 Nov;20(2):285-99. doi: 10.1006/prep.2000.1307.
A recombinant hexahistidine-tagged 18.5-kDa isoform of murine myelin basic protein has been characterized biochemically and immunogenically, by mass spectrometry, by circular dichroism under various conditions (in aqueous solution, with monosialoganglioside G(M1), and in 89% 2-propanol), and by transmission electron microscopy. The preparations of this protein indicated a high degree of purity and homogeneity, with no significant posttranslational modifications. Circular dichroic spectra showed that this preparation had the same degree of secondary structure as the natural bovine 18.5-kDa isoform of myelin basic protein. Incubation of the recombinant protein with lipid monolayers containing a nickel-chelating lipid resulted in the formation of fibrous assemblies that formed paracrystals of spacings 4.8 nm between fibers and 3-4 nm along them.
一种重组的带有六聚组氨酸标签的18.5 kDa小鼠髓鞘碱性蛋白亚型已通过质谱、在各种条件下(水溶液中、与单唾液酸神经节苷脂G(M1)一起、以及在89%异丙醇中)的圆二色性、以及透射电子显微镜进行了生化和免疫原性表征。该蛋白质的制剂显示出高度的纯度和均一性,没有明显的翻译后修饰。圆二色光谱表明,该制剂与天然牛18.5 kDa髓鞘碱性蛋白亚型具有相同程度的二级结构。将重组蛋白与含有镍螯合脂质的脂质单层一起孵育,导致形成纤维状聚集体,这些聚集体形成了纤维间距为4.8 nm且沿纤维方向为3 - 4 nm的准晶体。