Fanous Ali, Weiland Florian, Lück Carsten, Görg Angelika, Friess Albrecht, Parlar Harun
Technical University of Munich, Department for Chemical-Technical Analysis, Research Center for Brewing and Food Quality, Weihenstephaner Steig 23, D-85354 Freising-Weihenstephan, Germany.
Chemosphere. 2007 Aug;69(1):25-31. doi: 10.1016/j.chemosphere.2007.04.075. Epub 2007 Jun 12.
The herbicide 2,4-dichlorophenoxy acetic acid (2,4-D) induces a wide spectrum of toxic responses in living organisms. In this study, we analyzed the stress-induced responses of Corynebacterium glutamicum cells on protein level upon treatment with 2,4-D. For this, growing C. glutamicum cells were exposed to sublethal concentrations of 2,4-D, and changes of the gene expression profiles in comparison to non-exposed organisms were analyzed by two-dimensional gel electrophoresis and mass spectrometry. 2,4-D induced the over-expression of at least six C. glutamicum proteins, four of which could be identified by MALDI-TOF-MS. One protein (Cg2521; long-chain acyl-CoA synthetase) was related to the energy metabolism, and two proteins were involved in cell envelope synthesis (Cg2410; glutamine-dependent amidotransferase, and Cg1672; glycosyltransferase). The last induced protein was the ABC type transport system (Cg2695, ATPase component). The newly observed proteins, except for the ABC transport system, were not in general stress-related proteins, but were specifically expressed upon 2,4-D exposure and, therefore, can be used as respective biomarkers. Moreover, since these proteins seem to play a pivotal role in the adaptation of the cell to 2,4-D, they may help to gain deeper insight into the damage mechanisms of 2,4-D induced in the living cell.
除草剂2,4-二氯苯氧乙酸(2,4-D)可在活生物体中引发广泛的毒性反应。在本研究中,我们分析了谷氨酸棒杆菌细胞在2,4-D处理后蛋白质水平上的应激诱导反应。为此,将生长中的谷氨酸棒杆菌细胞暴露于亚致死浓度的2,4-D,并通过二维凝胶电泳和质谱分析与未暴露生物体相比基因表达谱的变化。2,4-D诱导了至少六种谷氨酸棒杆菌蛋白质的过度表达,其中四种可通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)鉴定。一种蛋白质(Cg2521;长链酰基辅酶A合成酶)与能量代谢相关,两种蛋白质参与细胞壁合成(Cg2410;谷氨酰胺依赖性酰胺转移酶和Cg1672;糖基转移酶)。最后一种诱导蛋白是ABC型转运系统(Cg2695,ATP酶成分)。新观察到的蛋白质,除了ABC转运系统外,一般都不是应激相关蛋白,而是在2,4-D暴露时特异性表达的,因此可以用作各自的生物标志物。此外,由于这些蛋白质似乎在细胞对2,4-D的适应中起关键作用,它们可能有助于更深入地了解2,4-D在活细胞中诱导的损伤机制。