Chaves Inês, Pinheiro Carla, Paiva Jorge A P, Planchon Sébastien, Sergeant Kjell, Renaut Jenny, Graça José A, Costa Gonçalo, Coelho Ana V, Ricardo Cândido P Pinto
Instituto de Tecnologia Química e Biológica (Universidade Nova de Lisboa), Portugal.
Proteomics. 2009 Sep;9(17):4154-75. doi: 10.1002/pmic.200700649.
Proteins from potato (Solanum tuberosum L.) tuber slices, related to the wound-healing process, were separated by 2-DE and identified by an MS analysis in MS and MS/MS mode. Slicing triggered differentiation processes that lead to changes in metabolism, activation of defence and cell-wall reinforcement. Proteins related to storage, cell growth and division, cell structure, signal transduction, energy production, disease/defence mechanisms and secondary metabolism were detected. Image analysis of the 2-DE gels revealed a time-dependent change in the complexity of the polypeptide patterns. By microscopic observation the polyalyphatic domain of suberin was clearly visible by D4, indicating that a closing layer (primary suberisation) was formed by then. A PCA of the six sampling dates revealed two time phases, D0-D2 and D4-D8, with a border position between D2 and D4. Moreover, a PCA of differentially expressed proteins indicated the existence of a succession of proteomic events leading to wound-periderm reconstruction. Some late-expressed proteins (D6-D8), including a suberisation-associated anionic peroxidase, have also been identified in the native periderm. Despite this, protein patterns of D8 slices and native periderm were still different, suggesting that the processes of wound-periderm formation are extended in time and not fully equivalent. The information presented in this study gives clues for further work on wound healing-periderm formation processes.
通过双向电泳(2-DE)分离了马铃薯(Solanum tuberosum L.)块茎切片中与伤口愈合过程相关的蛋白质,并采用质谱(MS)及串联质谱(MS/MS)模式进行了鉴定。切片引发了分化过程,导致代谢变化、防御激活和细胞壁强化。检测到了与储存、细胞生长和分裂、细胞结构、信号转导、能量产生、疾病/防御机制以及次生代谢相关的蛋白质。对双向电泳凝胶的图像分析揭示了多肽图谱复杂性的时间依赖性变化。通过显微镜观察,在第4天(D4)时可见木栓质的多脂肪区域,表明此时已形成封闭层(初级木栓化)。对六个采样日期进行主成分分析(PCA)显示出两个时间阶段,即D0 - D2和D4 - D8,在D2和D4之间存在边界位置。此外,对差异表达蛋白质的主成分分析表明存在一系列导致伤口周皮重建的蛋白质组学事件。一些后期表达的蛋白质(D6 - D8),包括一种与木栓化相关的阴离子过氧化物酶,也在天然周皮中被鉴定出来。尽管如此,D8切片和天然周皮的蛋白质模式仍然不同,这表明伤口周皮形成过程在时间上是延长的,且并不完全等同。本研究中呈现的信息为进一步研究伤口愈合 - 周皮形成过程提供了线索。