Protein composition of wheat gluten polymer fractions determined by quantitative two-dimensional gel electrophoresis and tandem mass spectrometry.
Protein composition of wheat gluten polymer fractions determined by quantitative two-dimensional gel electrophoresis and tandem mass spectrometry.
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DOI：
10.1186/1477-5956-12-8
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发表时间：
2014-02-11
期刊：
影响因子：
2
通讯作者：
Altenbach SB
中科院分区：
文献类型：
--
作者：
Vensel WH;Tanaka CK;Altenbach SB
某些小麦面筋蛋白形成大的蛋白质聚合物，其仅在超声处理后在0.5%SDS中可提取。虽然这些聚合物在面粉中的量和面包制作质量之间有很强的关系，但这些聚合物的蛋白质组分尚未得到彻底的研究。从美国面包小麦Butte 86中提取面粉蛋白质，在0.5%SDS中使用两步程序超声处理和不超声处理。通过尺寸排阻色谱法（SEC）将蛋白质进一步分离成单体和聚合物级分，并通过定量二维凝胶电泳（2-DE）进行分析。当选择2-DE斑点中的蛋白质通过串联质谱（MS/MS）鉴定时，来自不同蛋白质级分的重叠斑点通常产生不同的鉴定。高分子量麦谷蛋白亚基（HMW-GS）和低分子量麦谷蛋白亚基（LMW-GS）主要分布在聚合体组分中，醇溶蛋白主要分布在单体组分中。例外的是含有奇数半胱氨酸残基的α、γ和ω麦醇溶蛋白。在所有组分中均检测到这些蛋白质，但其占SDS可浸提聚合物组分的比例最大。在聚合物组分中还发现了几种类型的非谷蛋白，包括丝氨酸蛋白酶抑制剂、小麦蛋白和球蛋白。所有三种类型的发现在SDS-可提取的聚合物馏分中的比例最大。这是第一个研究报告积累的麦醇溶蛋白含有奇数的半胱氨酸残基的SDS-可提取的谷蛋白聚合物级分，支持的假设，这些麦醇溶蛋白作为链终止剂的聚合物链。这些数据使人们有可能制定关于蛋白质组成如何影响聚合物的大小和结构的假设，并为未来的实验提供了基础，旨在确定环境如何影响谷蛋白聚合物的分布。此外，分析揭示了小麦面粉蛋白质组的复杂性，在评估定量2-DE数据时应考虑到额外的层。
Certain wheat gluten proteins form large protein polymers that are extractable in 0.5% SDS only after sonication. Although there is a strong relationship between the amounts of these polymers in the flour and bread-making quality, the protein components of these polymers have not been thoroughly investigated. Flour proteins from the US bread wheat Butte 86 were extracted in 0.5% SDS using a two-step procedure with and without sonication. Proteins were further separated by size exclusion chromatography (SEC) into monomeric and polymeric fractions and analyzed by quantitative two-dimensional gel electrophoresis (2-DE). When proteins in select 2-DE spots were identified by tandem mass spectrometry (MS/MS), overlapping spots from the different protein fractions often yielded different identifications. Most high-molecular-weight glutenin subunits (HMW-GS) and low-molecular-weight glutenin subunits (LMW-GS) partitioned into the polymer fractions, while most gliadins were found in the monomer fractions. The exceptions were alpha, gamma and omega gliadins containing odd numbers of cysteine residues. These proteins were detected in all fractions, but comprised the largest proportion of the SDS-extractable polymer fraction. Several types of non-gluten proteins also were found in the polymer fractions, including serpins, triticins and globulins. All three types were found in the largest proportions in the SDS-extractable polymer fraction. This is the first study to report the accumulation of gliadins containing odd numbers of cysteine residues in the SDS-extractable glutenin polymer fraction, supporting the hypothesis that these gliadins serve as chain terminators of the polymer chains. These data make it possible to formulate hypotheses about how protein composition influences polymer size and structure and provide a foundation for future experiments aimed at determining how environment affects glutenin polymer distribution. In addition, the analysis revealed additional layers of complexity to the wheat flour proteome that should be considered when evaluating quantitative 2-DE data.
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影响因子：
3.8
作者：
Masci, S;Egorov, TA;Lafiandra, D
通讯作者：
Lafiandra, D
影响因子：
2.4
作者：
Cinco-Moroyoqui, Francisco J.;MacRitchie, Finlay
通讯作者：
MacRitchie, Finlay
影响因子：
2
作者：
Hurkman WJ;Tanaka CK;Vensel WH;Thilmony R;Altenbach SB
通讯作者：
Altenbach SB
影响因子：
3.8
作者：
Altenbach, SB;DuPont, FM;Lieu, D
通讯作者：
Lieu, D
影响因子：
3.8
作者：
GUPTA, RB;KHAN, K;MACRITCHIE, F
通讯作者：
MACRITCHIE, F