Heat-Treatment-Responsive Proteins in Different Developmental Stages of Tomato Pollen Detected by Targeted Mass Accuracy Precursor Alignment (tMAPA)

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• 作者：Palak Chaturvedi ; Hannes Doerfler ; Sridharan Jegadeesan ; Arindam Ghatak ; Etan Pressman ; Maria Angeles Castillejo ; Stefanie Wienkoop ; Volker Egelhofer ; Nurit Firon ; Wolfram Weckwerth
• 刊名：Journal of Proteome Research
• 出版年：2015
• 出版时间：November 6, 2015
• 年：2015
• 卷：14
• 期：11
• 页码：4463-4471
• 全文大小：467K
• ISSN：1535-3907

文摘

Recently, we have developed a quantitative shotgun proteomics strategy called mass accuracy precursor alignment (MAPA). The MAPA algorithm uses high mass accuracy to bin mass-to-charge (m/z) ratios of precursor ions from LC鈥揗S analyses, determines their intensities, and extracts a quantitative sample versus m/z ratio data alignment matrix from a multitude of samples. Here, we introduce a novel feature of this algorithm that allows the extraction and alignment of proteotypic peptide precursor ions or any other target peptide from complex shotgun proteomics data for accurate quantification of unique proteins. This strategy circumvents the problem of confusing the quantification of proteins due to indistinguishable protein isoforms by a typical shotgun proteomics approach. We applied this strategy to a comparison of control and heat-treated tomato pollen grains at two developmental stages, post-meiotic and mature. Pollen is a temperature-sensitive tissue involved in the reproductive cycle of plants and plays a major role in fruit setting and yield. By LC鈥揗S-based shotgun proteomics, we identified more than 2000 proteins in total for all different tissues. By applying the targeted MAPA data-processing strategy, 51 unique proteins were identified as heat-treatment-responsive protein candidates. The potential function of the identified candidates in a specific developmental stage is discussed.