| Combining Phytate/Ca~（2＋） Fractionation with Trichloroacetic Acid/Acetone Precipitation Improved Separation of Low-Abundant Proteins of Wheat （Triticum aestivum L.） Leaf for Proteomic Analysis |
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| 作者姓名： | Muhammad A R F Sultan LIU Hui CHENG Yu-Feng ZHANG Pei-pei ZHAO Hui-xian |
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| 作者单位： | State Key Laboratory of Crop Stress Biology for Arid Areas, Ministry of Science and Technology/College of Life Sciences, Northwest A＆F University |
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| 基金项目： | supported by the National Natural Science Foundation of China （30871578）; the Key Project of National Plant Transgenic Genes of China（2008ZX08002004,2011ZX08002004） |
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| 摘 要： | Proteomic assessment of low-abundance leaf proteins is hindered by the large quantity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) present within plant leaf tissues. In the present study, total proteins were extracted from wheat (Triticum aestivum L.) leaves by a conventional trichloroacetic acid (TCA)/acetone method and a protocol first developed in this work. Phytate/Ca2+ fractionation and TCA/acetone precipitation were combined to design an improved TCA/acetone method. The extracted proteins were analysed by two-dimensional gel electrophoresis (2-DE). The resulting 2-DE images were compared to reveal major differences. The results showed that large quantities of Rubisco were deleted from wheat leaf proteins prepared by the improved method. As many as (758±4) protein spots were detected from 2-DE images of protein extracts obtained by the improved method, 130 more than those detected by the TCA/acetone method. Further analysis indicated that more protein spots could be detected at regions of pI 4.00-4.99 and 6.50-7.00 in the improved method-based 2-DE images. Our findings indicated that the improved method is an efficient protein preparation protocol for separating low-abundance proteins in wheat leaf tissues by 2-DE analysis. The proposed protocol is simple, fast, inexpensive and also applicable to protein preparations of other plants.
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| 关 键 词： | Triticum aestivum L. Rubisco low-abundance protein phytate/Ca2+ two-dimensional gel electrophoresis plant leaf proteomics |
Combining Phytate/Ca~（2＋） Fractionation with Trichloroacetic Acid/Acetone Precipitation Improved Separation of Low-Abundant Proteins of Wheat （Triticum aestivum L.） Leaf for Proteomic Analysis |
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| Muhammad,A,R,F,Sultan LIU,Hui CHENG,Yu-Feng ZHANG,Pei-pei ZHAO,Hui-xian.Combining Phytate/Ca~（2＋） Fractionation with Trichloroacetic Acid/Acetone Precipitation Improved Separation of Low-Abundant Proteins of Wheat （Triticum aestivum L.） Leaf for Proteomic Analysis[J].Agricultural Sciences in China,2013(7):1123-1129. |
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| Authors: | Agricultural Sciences in China |
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| Institution: | Key Laboratory of Crop Stress Biology for Arid Areas, Ministry of Science and Technology/College of Life Sciences, Northwest A&F University, Yangling 712100, P.R.China |
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| Abstract: | |
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| Keywords: | Triticum aestivum L Rubisco low-abundance protein phytate/Ca2＋ two-dimensional gel electrophoresis plant leaf proteomics |
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