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首页> 外文期刊>Journal of Agricultural and Food Chemistry >Qualitative and quantitative evaluation of protein extraction protocols for apple and strawberry fruit suitable for two-dimensional electrophoresis and mass spectrometry analysis.
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Qualitative and quantitative evaluation of protein extraction protocols for apple and strawberry fruit suitable for two-dimensional electrophoresis and mass spectrometry analysis.

机译：苹果和草莓果实蛋白质提取方案的定性和定量评估，适用于二维电泳和质谱分析。

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A modified phenol-based protocol and a phenol-free protocol that involves hot SDS extraction followed by TCA precipitation in acetone were qualitatively and quantitatively compared and evaluated on apple peel and strawberry fruit. The phenol protocol resulted in significantly higher protein yields of 2.35 +/- 0.1 and 0.46 +/- 0.06 mg/g of FW from apple and strawberry fruit, respectively, compared to the SDS protocol, which produced 0.74 +/- 0.1 and 0.27 +/- 0.02 mg/g of FW, respectively. 2-DE analysis of apple protein extracts revealed 1422 protein spots associated with the phenol protocol and 849 spots associated with the SDS protocol. Of these, 761 were present only in phenol gels, whereas 23 were exclusive to SDS samples. For strawberry, SDS extraction produced poor-quality spots with a high degree of streaking, indicating possible contamination. The application of a cleanup procedure resulted in a purified protein extract with high-quality spots. 2-DE analysis of strawberry protein extracts revealed 1368 spots for the phenol protocol and 956 spots for the SDS protocol accompanied by the cleanup procedure. Of these, 599 spots were present only in phenol gels, whereas 109 were present only in SDS samples. Spots from each fruit tissue and extraction procedure were selected, and a total of 26 were identified by LC-MS/MS. Overall, this study demonstrates the complexity of protein extraction of fruit tissues and suggests that a phenol-based protein extraction protocol should be used as a standard procedure for recalcitrant fruit tissues, whereas a SDS protocol with or without a cleanup procedure may be used as an alternative protocol.
机译：在苹果皮和草莓果实上定性和定量地比较和评估了改进的基于酚的方案和涉及热SDS提取然后在丙酮中TCA沉淀的无酚方案。与SDS方案产生的0.74 +/- 0.1和0.27 +的SDS方案相比，酚方案产生的苹果和草莓果实的FW分别分别高出2.35 +/- 0.1和0.46 +/- 0.06 mg / g的蛋白质产量。分别为0.02 mg / g FW。苹果蛋白质提取物的2-DE分析显示，与酚规程相关的1422个蛋白斑点和与SDS规程相关的849个斑点。其中761种仅存在于酚凝胶中，而23种仅在SDS样品中存在。对于草莓，SDS提取产生的斑点质量很差，且条纹程度很高，表明可能存在污染。应用清除程序可得到具有高质量斑点的纯化蛋白提取物。草莓蛋白提取物的2-DE分析显示，苯酚操作规程为1368个斑点，SDS操作规程为956个斑点，并伴有清除步骤。其中599个斑点仅存在于酚凝胶中，而109个斑点仅存在于SDS样品中。从每个水果组织和提取程序中选择斑点，并通过LC-MS / MS鉴定出总共26个斑点。总的来说，这项研究证明了水果组织蛋白质提取的复杂性，并建议应将基于酚的蛋白质提取方案用作顽固性水果组织的标准程序，而可以将采用或不采用清除程序的SDS方案用作替代协议。

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