Gerber Isak B, Laukens Kris, De Vijlder Thomas, Witters Erwin, Dubery Ian A
Department of Biochemistry, University of Johannesburg, P.O. Box 524, Auckland Park, 2006, South Africa.
Biochim Biophys Acta. 2008 Nov;1784(11):1750-62. doi: 10.1016/j.bbapap.2008.06.012. Epub 2008 Jun 25.
Plants constantly monitor for pathogen challenge and utilize a diverse array of adaptive defense mechanisms, including differential protein regulation, during pathogen attack. A proteomic analysis of Nicotiana tabacum BY-2 cells was performed in order to investigate the dynamic changes following perception of bacterial lipopolysaccharides. A multiplexed proteome analysis, employing two-dimensional difference-in-gel-electrophoresis with CyDye DIGE fluors, as well as Ruthenium II tris (bathophenanthroline disulfonate) fluorescence staining and Pro-Q Diamond phosphoprotein-specific gel staining, monitored over 1500 proteins and resulted in the identification of 88 differentially regulated proteins and phosphoproteins responsive to LPS(B.cep.)-elicitation. Functional clustering of the proteins both at the level of their abundance and phosphorylation status, revealed 9 proteins involved in transport, ion homeostasis and signal transduction. A large number of responsive proteins were found to be involved in metabolism- and energy-related processes (36), representing various metabolic pathways. Another abundant category corresponded to proteins classified as molecular chaperones and involved in protein destination/targeting (12). Other categories of proteins found to be LPS(B.cep.)-responsive and differentially regulated include cell structure- and cytoskeletal rearrangement proteins (8) and proteins involved in transcription and translation as well as degradation (11). The results indicate that LPS(B.cep.) induces metabolic reprogramming and changes in cellular activities supporting protein synthesis, -folding, vesicle trafficking and secretion; accompanied by changes to the cytoskeleton and proteosome function. Many of the identified proteins are known to be interconnected at various levels through a complex web of activation/deactivation, complex formation, protein-protein interactions, and chaperoning reactions. The presented data offers novel insights and further evidence for the biochemical action of LPS(B.cep.) as a resistance elicitor, a pathogen-associated molecular pattern molecule and triggering agent of defense responses associated with innate immunity.
植物不断监测病原体的挑战，并在病原体攻击期间利用多种适应性防御机制，包括差异蛋白质调节。为了研究烟草BY-2细胞在感知细菌脂多糖后的动态变化，进行了蛋白质组学分析。采用二维差异凝胶电泳结合CyDye DIGE荧光染料、钌（II）三（邻菲罗啉二磺酸盐）荧光染色和Pro-Q Diamond磷蛋白特异性凝胶染色的多重蛋白质组分析，监测了1500多种蛋白质，鉴定出88种对LPS（B.cep.）诱导有差异调节的蛋白质和磷蛋白。对蛋白质在丰度和磷酸化状态水平上的功能聚类，揭示了9种参与运输、离子稳态和信号转导的蛋白质。发现大量响应蛋白参与代谢和能量相关过程（36种），代表了各种代谢途径。另一个丰富的类别对应于归类为分子伴侣并参与蛋白质定位/靶向的蛋白质（12种）。发现对LPS（B.cep.）有响应并差异调节的其他蛋白质类别包括细胞结构和细胞骨架重排蛋白（8种）以及参与转录、翻译和降解的蛋白质（11种）。结果表明，LPS（B.cep.）诱导代谢重编程和细胞活动变化，支持蛋白质合成、折叠、囊泡运输和分泌；同时伴随着细胞骨架和蛋白酶体功能的变化。许多已鉴定的蛋白质已知通过激活/失活、复合物形成、蛋白质-蛋白质相互作用和伴侣反应的复杂网络在各个层面相互连接。所呈现的数据为LPS（B.cep.）作为抗性激发子、病原体相关分子模式分子和与先天免疫相关的防御反应触发剂的生化作用提供了新的见解和进一步的证据。
