Schmid Roland, Bernhardt Jörg, Antelmann Haike, Völker Andrea, Mach Hiltraut, Völker Uwe, Hecker Michael
Universität Osnabrük, Abteilung für Mikrobiologie, 49076 Osnabrück, Germany.
Institut für Mikrobiologie und Molekularbiologie, Ernst-Moritz-Arndt-Universität Greifswald, 17487 Greifswald, Friedrich-Ludwig-Jahn-StraBe 15, Germany.
Microbiology (Reading). 1997 Mar;143 ( Pt 3):991-998. doi: 10.1099/00221287-143-3-991.
Twenty-three of the most prominent spots which are visible on two-dimensional (2-D) protein gels of Bacillus subtilis crude extracts were selected as marker spots for the construction of a 2-D protein index. N-terminal sequencing of the corresponding proteins resulted in the identification of enzymes involved in glycolysis, TCA cycle, pentose phosphate cycle, amino acid metabolism, nucleotide biosynthesis and translation. Using computer analysis of the 2-D protein gels, most of these metabolic enzymes were found to be synthesized at a reduced rate after different stresses and glucose starvation. Such an approach permits a rapid and global evaluation of the regulation of different branches of metabolism in response to various physiological conditions.
从枯草芽孢杆菌粗提物的二维(2-D)蛋白质凝胶上可见的23个最显著斑点中,选择作为构建二维蛋白质索引的标记斑点。对相应蛋白质进行N端测序,鉴定出参与糖酵解、三羧酸循环、磷酸戊糖循环、氨基酸代谢、核苷酸生物合成和翻译的酶。通过对二维蛋白质凝胶的计算机分析,发现这些代谢酶中的大多数在不同应激和葡萄糖饥饿后合成速率降低。这种方法可以快速、全面地评估不同代谢分支在各种生理条件下的调节情况。