Rubikas J, Androsiuniene D, Chestukhina G, Smirnova T, Kapitonova O, Stepanov V
Institute of Biochemistry, Academy of Sciences of the Lithuanian SSR, Vilnius, USSR.
J Bacteriol. 1987 Nov;169(11):5258-62. doi: 10.1128/jb.169.11.5258-5262.1987.
Electron microscopic investigation of ultrathin sections of Bacillus subtilis Cgr4 cells revealed the presence of crystal-like inclusions which were formed of spheric homogeneous subunits. The frequency of cells with a crystal-like inclusion in the culture approached 1%. The appearance of the crystal protein in cells coincided in time with spore morphogenesis. However, the process of crystal protein formation and sporulation are two alternatives: the cells either form the crystal protein or continue spore morphogenesis. Fractionation of cells in the stationary growth phase on a Percoll density gradient showed that the cells containing the crystal protein accumulated in the fraction corresponding to a 1.14-g/ml Percoll density. The cells were disintegrated by sonication, and alkaline-extracted proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. After sodium dodecyl sulfate-gel electrophoresis, the fraction enriched with crystal-containing cells showed practically a single band with a molecular weight of 47,000 that corresponded to the crystal-forming protein. The antigenic features and amino acid composition indicated certain similarities between the crystal-forming protein in B. subtilis Cgr4 cells and the spore coat protein.
对枯草芽孢杆菌Cgr4细胞超薄切片的电子显微镜研究显示,存在由球形均匀亚基构成的晶体状内含物。培养物中含有晶体状内含物的细胞频率接近1%。细胞中晶体蛋白的出现时间与孢子形态发生一致。然而,晶体蛋白形成过程和孢子形成是两种相互排斥的过程:细胞要么形成晶体蛋白,要么继续进行孢子形态发生。在Percoll密度梯度上对稳定生长期的细胞进行分级分离,结果表明,含有晶体蛋白的细胞聚集在对应于1.14 g/ml Percoll密度的级分中。通过超声破碎细胞,并对碱性提取的蛋白质进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析。十二烷基硫酸钠凝胶电泳后,富含含晶体细胞的级分显示出一条几乎单一的条带,其分子量为47,000,对应于形成晶体蛋白。抗原特性和氨基酸组成表明,枯草芽孢杆菌Cgr4细胞中的晶体形成蛋白与孢子外壳蛋白之间存在某些相似性。