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利用一维电泳和二维电泳分离结合串联质谱法解析莲(Nelumbo nucifera Gaertn.)种子胚乳蛋白质组

Unraveling the seed endosperm proteome of the lotus (Nelumbo nucifera Gaertn.) utilizing 1DE and 2DE separation in conjunction with tandem mass spectrometry.

作者信息

Moro Carlo F, Fukao Yoichiro, Shibato Junko, Rakwal Randeep, Timperio Anna Maria, Zolla Lello, Agrawal Ganesh Kumar, Shioda Seiji, Kouzuma Yoshiaki, Yonekura Masami

机构信息

Laboratory of Molecular Food Functionality, College of Agriculture, Ami, Ibaraki, Japan.

Plant Global Educational Project, Nara Institute of Science and Technology, Ikoma, Japan.

出版信息

Proteomics. 2015 May;15(10):1717-35. doi: 10.1002/pmic.201400406. Epub 2015 Feb 12.

Abstract

Nelumbo nucifera (Gaertn.) or lotus, is an aquatic plant native to India, and presently consumed as food mainly in China and Japan. Lotus is also widely used in Indian and Chinese traditional medicine. Extracts from different parts of the lotus plant have been reported to show diverse biological activities-antioxidant, free radical scavenging, anti-inflammatory, and immunomodulatory. Despite this, little work has been done in isolating and identifying proteins responsible for these activities, or yet importantly to establish a lotus proteome. The aim of our group is to develop a proteome catalog of the lotus plant, starting with its seed, the nutrient rich food source. In this present study, the seed endosperm-most abundant in proteins, and main nutrient storage tissue-was targeted for protein extraction by testing five different extraction protocols, followed by their proteomic analyses using complementary 1DE and 2DE approaches in conjunction with MS/MS. The inventory of 66 nonredundant proteins obtained by 1DE-MS and the 30 obtained by 2DE-MS provides the first catalog of the lotus seed endosperm, where the most abundant protein functions were in categories of metabolic activities related to carbohydrate metabolism and nutrient storage.

摘要

莲(Nelumbo nucifera (Gaertn.)),即荷花,是一种原产于印度的水生植物,目前主要在中国和日本作为食物食用。荷花在印度和中国传统医学中也有广泛应用。据报道,莲植物不同部位的提取物具有多种生物活性——抗氧化、清除自由基、抗炎和免疫调节。尽管如此,在分离和鉴定负责这些活性的蛋白质方面,或者更重要的是建立莲蛋白质组方面,所做的工作很少。我们小组的目标是从莲子(营养丰富的食物来源)开始,开发莲植物的蛋白质组目录。在本研究中,通过测试五种不同的提取方案,以蛋白质含量最丰富且主要的营养储存组织——种子胚乳为目标进行蛋白质提取,随后使用互补的一维电泳(1DE)和二维电泳(2DE)方法结合串联质谱(MS/MS)对其进行蛋白质组分析。通过一维电泳 - 质谱(1DE-MS)获得的66种非冗余蛋白质清单以及通过二维电泳 - 质谱(2DE-MS)获得的30种蛋白质清单提供了莲种子胚乳的首个目录,其中最丰富的蛋白质功能属于与碳水化合物代谢和营养储存相关的代谢活动类别。

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