Characterization of HMW glutenin subunits in common wheat and related species by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS)

文献类型： 外文期刊

第一作者： Zhang, Qian

作者： Zhang, Qian;Dong, Yanmin;An, Xueli;Wang, Aili;Zhang, Yanzhen;Li, Xiaohui;Gao, Liyan;Yan, Yueming;Xia, Xianchun;He, Zhonghu;Xia, Xianchun;He, Zhonghu

作者机构：

关键词： HMW-GS;molecular weight;mass spectrometry;PTM

期刊名称：JOURNAL OF CEREAL SCIENCE （ 影响因子：3.616； 五年影响因子：3.891 ）

ISSN： 0733-5210

年卷期： 2008 年 47 卷 2 期

页码：

收录情况： SCI

摘要： The sample preparation method of high molecular weight glutenin subunits (HMW-GS) for matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis, without a separation step, by high-performance liquid chromatography (HPLC) was established in this study. Three major factors influencing mass spectra-the ratio of components of the solvent, the resolving time, and the sample volume-were optimized using HMW-GS mixtures extracted from Chinese cultivar Jing 411. The results showed that the optimized method for sample preparation was to resolve HMW-GS from 20 mg in an hour with 50 mu 1 solvent of 0.4% TFA, 30.0% ACN and 69.6% H2O. The stable mass spectra and accurate molecular weights of 16 major HMW glutenin subunits from common wheat and related species were obtained using the optimized MALDI-TOF-MS method. Seven subunits, where each was from 2-5 cultivars, showed very similar molecular weights. The determined molecular weights of I I subunits were close to those calculated from their coding sequences. In addition, no positive reaction between HMW-GS and GelCode((R)) Glycoprotein Staining Reagent was observed. These results suggested that HMW-GS lack extensive post-translational modifications (PTMs), but low levels of glycosylation or phosphorylation present in some subunits cannot be ruled out. Because of its ability to obtain a rapid, complete and precise profile of HMW glutenin subunits without purifying procedures, MALDI-TOF-MS is expected to be a powerful technique for structural and functional studies of HMW glutenin subunits as well as other cereal proteins. (c) 2007 Elsevier Ltd. All rights reserved.

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