A Combined Method of Protein Extraction from Unorthodox Plant Samples for Proteomics

IF 0.5 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Current Proteomics Pub Date : 2020-12-09 DOI:10.2174/1570164618999201209221340

C. Yılmaz, M. Işcan

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Abstract

This study aimed to generate an improved method of protein extraction and purification from plant tissues containing very high amounts of phenolic compounds and other interfering biomolecules. Protein extraction at proteomic studies on some plant species including conifers is challenging and the yield and quality is unpredictable. Two popular protocols were combined with each other to construct a novel one with enhanced abilities to produce higher purity of samples compatible for high precision molecular systems and analysis. The new method was compared with the other two for their efficiencies in classical SDS-PAGE, 2-DE and capillary chromatography applications. All three methods were comparable in SDS-PAGE procedure; however, only the new method created acceptable gel images in 2-DE. Bioanalyzer results, also, demonstrated that the new method provided protein samples pure enough to be used in capillary chromatography with 2 times more peaks in electropherograms with lower noise and higher total relative protein concentrations closest to the applied amount. The new combined method is a successful alternative for plant proteomicists with higher yield and quality of proteins from recalcitrant tissues. The new method could be preferred, especially, for high-tech, sensitive proteomic analysis.

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一种从非正统植物样品中提取蛋白质的组合方法

本研究旨在建立一种从含有大量酚类化合物和其他干扰生物分子的植物组织中提取和纯化蛋白质的改进方法。在包括针叶树在内的一些植物的蛋白质组学研究中，蛋白质的提取具有挑战性，其产量和质量难以预测。将两种流行的方案相互结合，构建了一种新的方案，该方案具有增强的能力，可以产生更高纯度的样品，与高精度分子系统和分析兼容。将该方法与其他两种方法在经典的SDS-PAGE、2-DE和毛细管色谱中的效率进行了比较。3种方法在SDS-PAGE程序中具有可比性;然而，只有新方法在2-DE中创建了可接受的凝胶图像。生物分析仪的结果也表明，新方法提供的蛋白质样品纯度足以用于毛细管色谱，其电泳峰数增加了2倍，噪声更低，总相对蛋白质浓度更高，最接近施药量。该方法对植物蛋白质组学家来说是一种成功的替代方法，可以从顽固组织中获得更高的蛋白质产量和质量。这种新方法尤其适用于高科技、敏感的蛋白质组学分析。

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来源期刊

Current Proteomics BIOCHEMICAL RESEARCH METHODS-BIOCHEMISTRY & MOLECULAR BIOLOGY

CiteScore

1.60

自引率

0.00%

发文量

审稿时长

>0 weeks

期刊介绍： Research in the emerging field of proteomics is growing at an extremely rapid rate. The principal aim of Current Proteomics is to publish well-timed in-depth/mini review articles in this fast-expanding area on topics relevant and significant to the development of proteomics. Current Proteomics is an essential journal for everyone involved in proteomics and related fields in both academia and industry. Current Proteomics publishes in-depth/mini review articles in all aspects of the fast-expanding field of proteomics. All areas of proteomics are covered together with the methodology, software, databases, technological advances and applications of proteomics, including functional proteomics. Diverse technologies covered include but are not limited to: Protein separation and characterization techniques 2-D gel electrophoresis and image analysis Techniques for protein expression profiling including mass spectrometry-based methods and algorithms for correlative database searching Determination of co-translational and post- translational modification of proteins Protein/peptide microarrays Biomolecular interaction analysis Analysis of protein complexes Yeast two-hybrid projects Protein-protein interaction (protein interactome) pathways and cell signaling networks Systems biology Proteome informatics (bioinformatics) Knowledge integration and management tools High-throughput protein structural studies (using mass spectrometry, nuclear magnetic resonance and X-ray crystallography) High-throughput computational methods for protein 3-D structure as well as function determination Robotics, nanotechnology, and microfluidics.