Department of Plant Biology, University of Minnesota, St. Paul, MN 55108, USA.
Plant J. 2010 Aug;63(4):680-95. doi: 10.1111/j.1365-313X.2010.04266.x.
Rapid environmental responses in plants rely on endogenous signaling mechanisms, which in many cases are mediated by changes in protein turnover rates. It is therefore necessary to develop methods for measuring protein dynamics that monitor large sets of plant proteins to begin to apply a systems biology approach to the study of plant behavior. The use of stable isotope labeling strategies that are adaptable to proteomic methods is particularly attractive for this purpose. Here, we explore one example of such methods that is particularly suitable for plants at the seedling stage, where measurement of amino acid and protein turnover rates is accomplished using a heavy water labeling strategy. The method is backed by microarray evaluation to define its feasibility for specific experimental approaches, and the CULLIN-ASSOCIATED AND NEDDYLATION DISSOCIATED 1 (CAND1) and TRANSPORT INHIBITOR RESPONSE 1 (TIR1) proteins are used to illustrate the potential utility in understanding hormonal signaling regulation. These studies provide insight not only into the potential utility of the method, but also address possible areas of concern regarding the use of heavy water labeling during plant growth. These considerations suggest a prescription for specific experimental designs that minimize interference resulting from the induction of treatment-specific gene expression in the results obtained.
植物的快速环境响应依赖于内源性信号机制，而在许多情况下，这些信号机制是通过蛋白质周转率的变化来介导的。因此，有必要开发用于测量蛋白质动态的方法，以监测大量植物蛋白质，从而开始将系统生物学方法应用于植物行为的研究。使用可适应蛋白质组学方法的稳定同位素标记策略在这方面特别有吸引力。在这里，我们探讨了一种特别适用于幼苗阶段植物的方法示例，该方法使用重水标记策略来测量氨基酸和蛋白质周转率。该方法通过微阵列评估来支持其特定实验方法的可行性，并用 CULLIN-ASSOCIATED AND NEDDYLATION DISSOCIATED 1（CAND1）和 TRANSPORT INHIBITOR RESPONSE 1（TIR1）蛋白来说明其在理解激素信号转导调节方面的潜在应用。这些研究不仅提供了对该方法潜在应用的深入了解，还解决了在植物生长过程中使用重水标记可能存在的一些关注问题。这些考虑因素为具体的实验设计提供了建议，这些设计可最大限度地减少由于诱导特定于处理的基因表达而导致的结果中的干扰。
