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人参叶对高光响应的蛋白质组分析:电喷雾电离四极杆-飞行时间质谱法及表达序列标签数据的应用

Proteome analysis of the responses of Panax ginseng C. A. Meyer leaves to high light: use of electrospray ionization quadrupole-time of flight mass spectrometry and expressed sequence tag data.

作者信息

Nam Myung Hee, Heo Eun Joo, Kim Jin Young, Kim Seung Il, Kwon Kyung-Hoon, Seo Jong Bok, Kwon Ohoak, Yoo Jong Shin, Park Young Mok

机构信息

Seoul Branch, Korea Basic Science Institute, Seoul, Republic of South Korea.

出版信息

Proteomics. 2003 Dec;3(12):2351-67. doi: 10.1002/pmic.200300509.

DOI:10.1002/pmic.200300509
PMID:14673786
Abstract

We performed comparative proteomic analyses in order to understand the physiological responses of ginseng (Panax ginseng C. A. Meyer) to high light (HL). As a first step, we analyzed the proteins expressed in ginseng leaves. Proteins extracted from leaves were separated by two-dimensional polyacrylamide gel electrophoresis. Protein spots were identified by tandem mass spectra analysis using electrospray ionization quadrupole-time of flight mass spectrometry (ESI Q-TOF MS). We used a ginseng expressed sequence tag (EST) database as well as a nonredundant protein database from NCBI to identify proteins. Eighty-one proteins were identified using the nr protein database, 51 of which were also verified from the ginseng EST database. An additional 66 proteins were identified only from the ginseng EST database. Proteins that function in energy metabolism, protein stabilization, and protection against oxidative stress were abundant. To understand the light responses of ginseng leaves, we studied time dependent changes in expressed proteins produced by 0-4 h of HL exposure. Six HL-responsive proteins were identified: three proteins were up-regulated (cytosolic small heat-shock protein, cytosolic ascorbate peroxidase, and putative major latex-like protein) and three proteins were down-regulated (Rieske Fe/S protein, putative 3-beta hydroxysteroid dehydrogenase/isomerase-like protein, and oxygen-evolving enhancer-like protein). Our results show that the ginseng EST database combined with ESI Q-TOF MS analysis can be used to identify ginseng proteins and to elucidate the protective mechanism of ginseng against HL induced damage.

摘要

为了解人参(Panax ginseng C. A. Meyer)对强光(HL)的生理反应,我们进行了比较蛋白质组学分析。第一步,我们分析了人参叶片中表达的蛋白质。从叶片中提取的蛋白质通过二维聚丙烯酰胺凝胶电泳进行分离。使用电喷雾电离四极杆-飞行时间质谱(ESI Q-TOF MS)通过串联质谱分析鉴定蛋白质斑点。我们使用人参表达序列标签(EST)数据库以及来自NCBI的非冗余蛋白质数据库来鉴定蛋白质。使用nr蛋白质数据库鉴定出81种蛋白质,其中51种也从人参EST数据库中得到验证。另外66种蛋白质仅从人参EST数据库中鉴定出来。在能量代谢、蛋白质稳定和抗氧化应激方面发挥作用的蛋白质含量丰富。为了解人参叶片的光反应,我们研究了0-4小时HL处理后表达蛋白质的时间依赖性变化。鉴定出6种HL反应蛋白:3种蛋白上调(胞质小热休克蛋白、胞质抗坏血酸过氧化物酶和假定的主要乳胶样蛋白),3种蛋白下调( Rieske Fe/S蛋白、假定的3-β羟类固醇脱氢酶/异构酶样蛋白和放氧增强子样蛋白)。我们的结果表明,人参EST数据库与ESI Q-TOF MS分析相结合可用于鉴定人参蛋白质,并阐明人参对HL诱导损伤的保护机制。

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