Hansmeier Nicole, Chao Tzu-Chiao, Pühler Alfred, Tauch Andreas, Kalinowski Jörn
Lehrstuhl für Genetik, Fakultät für Biologie, Universität Bielefeld, Universitätsstrasse 25, D-33615 Bielefeld, Germany.
Proteomics. 2006 Jan;6(1):233-50. doi: 10.1002/pmic.200500144.
Reference maps of the cytosolic, cell surface and extracellular proteome fractions of the amino acid-producing soil bacterium Corynebacterium efficiens YS-314 were established. The analysis window covers a pI range from 3 to 7 along with a molecular mass range from 10 to 130 kDa. After second-dimensional separation on SDS-PAGE and Coomassie staining, computational analysis detected 635 protein spots in the cytosolic proteome fraction, whereas 76 and 102 spots were detected in the cell surface and extracellular proteomes, respectively. By means of MALDI-TOF-MS and tryptic peptide mass fingerprinting, 164 cytosolic proteins, 49 proteins of the cell surface and 89 extracellular protein spots were identified, representing in total 177 different proteins. Additionally, reference maps of the three cellular proteome fractions of the close phylogenetic relative Corynebacterium glutamicum ATCC 13032 were generated and used for comparative proteomics. Classification according to the Clusters of Orthologous Groups of proteins scheme and abundance analysis of the identified proteins revealed species-specific differences. The high abundance of molecular chaperones and amino acid biosynthesis enzymes in C. efficiens points to environmental adaptations of this recently discovered amino acid-producing bacterium.
建立了产氨基酸土壤细菌高效棒状杆菌YS-314的胞质、细胞表面和细胞外蛋白质组级分的参考图谱。分析窗口涵盖了3至7的pI范围以及10至130 kDa的分子量范围。在SDS-PAGE上进行二维分离并经考马斯亮蓝染色后,计算分析在胞质蛋白质组级分中检测到635个蛋白点,而在细胞表面和细胞外蛋白质组中分别检测到76个和102个蛋白点。通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)和胰蛋白酶肽质量指纹图谱,鉴定出164个胞质蛋白、49个细胞表面蛋白和89个细胞外蛋白点,总共代表177种不同的蛋白质。此外,还生成了近缘系统发育相关菌谷氨酸棒状杆菌ATCC 13032的三个细胞蛋白质组级分的参考图谱,并将其用于比较蛋白质组学研究。根据蛋白质直系同源簇(COG)分类方案对鉴定出的蛋白质进行分类并进行丰度分析,揭示了物种特异性差异。高效棒状杆菌中分子伴侣和氨基酸生物合成酶的高丰度表明这种最近发现的产氨基酸细菌对环境的适应性。