Smith R D, Salyers A A
J Bacteriol. 1981 Jan;145(1):8-13. doi: 10.1128/jb.145.1.8-13.1981.
L-[4,5-3H]- or L-[U-14C]leucine was incorporated by Bacteroides thetaiotaomicron into acid-precipitable material even when the bacteria were treated with concentrations of tetracycline high enough to prevent growth. Similar results were obtained when L-[2,3,4-3H]valine or L-[4,5-3H]isoleucine was used instead of leucine. In bacteria which had been treated with tetracycline, the acid-precipitable label was not solubilized by treatment with protease, lysozyme, or deoxyribonuclease. However, virtually all of the label was extractable with chloroform-methanol, indicating that the label had been incorporated into membrane lipids. Since L-[1-14C]leucine was not incorporated into lipids, leucine was probably decarboxylated before incorporation. When a chloroform extract from bacteria which had been labeled with both [32P]phosphate and [3H]leucine was resolved into component phospholipids by two-dimensional thin-layer chromatography, 3H was incorporated into all of the phospholipids. When these phospholipids were deacylated, the 3H from leucine was associated with released fatty acids rather than with the head groups. Thus, it appears that B. thetaiotaomicron can utilize leucine and similar amino acids not only by incorporating them into protein but also by incorporating portions of these amino acids into membrane phospholipids.
即使将拟杆菌用高浓度的四环素处理以阻止其生长,L-[4,5-³H]-或L-[U-¹⁴C]亮氨酸仍能被拟杆菌掺入酸沉淀物质中。当用L-[2,3,4-³H]缬氨酸或L-[4,5-³H]异亮氨酸代替亮氨酸时,也得到了类似的结果。在用四环素处理过的细菌中,酸沉淀标记物不会因蛋白酶、溶菌酶或脱氧核糖核酸酶的处理而溶解。然而,几乎所有的标记物都可用氯仿-甲醇提取,这表明标记物已掺入膜脂中。由于L-[1-¹⁴C]亮氨酸未掺入脂质中,亮氨酸可能在掺入前就已脱羧。当用二维薄层色谱法将用[³²P]磷酸盐和[³H]亮氨酸标记的细菌的氯仿提取物分离成组成磷脂时,³H被掺入所有的磷脂中。当这些磷脂被脱酰基时,来自亮氨酸的³H与释放的脂肪酸相关,而不是与头部基团相关。因此,看来拟杆菌不仅可以通过将亮氨酸和类似氨基酸掺入蛋白质中,还可以通过将这些氨基酸的部分掺入膜磷脂中来利用它们。