Analysis of expressed sequence tags from a single wheat cultivar facilitates interpretation of tandem mass spectrometry data and discrimination of gamma gliadin proteins that may play different functional roles in flour.
Analysis of expressed sequence tags from a single wheat cultivar facilitates interpretation of tandem mass spectrometry data and discrimination of gamma gliadin proteins that may play different functional roles in flour.
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DOI：
10.1186/1471-2229-10-7
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发表时间：
2010-01-11
影响因子：
5.3
通讯作者：
Dupont FM
中科院分区：
文献类型：
--
作者：
Altenbach SB;Vensel WH;Dupont FM
γ醇溶蛋白是一组复杂的蛋白质，与其他面筋蛋白一起决定小麦粉的功能特性。这些蛋白质含有异常高水平的谷氨酰胺和脯氨酸，并含有大面积的重复序列。虽然大多数γ-麦醇溶蛋白是含有8个保守的半胱氨酸残基的单体蛋白质，但有些含有额外的半胱氨酸残基，使它们能够与其他面筋蛋白连接成对面粉质量至关重要的大聚合物。区分γ醇溶蛋白的能力对于小麦粉品质的研究是重要的，因为具有相似序列的蛋白质可以对功能特性具有不同的影响。通过分析公开可用的表达序列标签（EST）数据来评估在小麦栽培品种Butte 86中表达的γ醇溶蛋白基因的互补。从153个Butte 86 EST中组装了11个重叠群。9个重叠群编码全长蛋白质，4个蛋白质含有9个半胱氨酸残基。只有一种编码的蛋白质与NCBI报道的序列完全匹配。来自四种不同的公开可用的EST组装体的重叠群编码的蛋白质与Butte 86 γ醇溶蛋白中的一些（但不是全部）完美匹配，并且相同蛋白质的互补对于每个组装体是不同的。一个专门的数据库，其中包括Butte 86 γ醇溶蛋白的序列构建用于通过串联质谱法（MS/MS）鉴定面粉蛋白。在试验性实验中，通过MS/MS区分对应于6个Butte 86 γ麦醇溶蛋白重叠群的蛋白质，包括含有额外半胱氨酸残基的蛋白质。另外两种蛋白质被鉴定为两种密切相关的Butte 86蛋白质之一，但不能明确区分。报道了对Butte 86 γ醇溶蛋白具有特异性的独特肽标签。包括在数据库中的品种特定的γ麦醇溶蛋白序列的数量和质量的肽鉴定，并增加这些γ麦醇溶蛋白的序列覆盖率的MS/MS。这种方法使得有可能区分密切相关的蛋白质，将个别蛋白质与特定基因的序列，并评估蛋白质组学数据在生物背景下，以更好地解决小麦面粉质量的问题。
The gamma gliadins are a complex group of proteins that together with other gluten proteins determine the functional properties of wheat flour. The proteins have unusually high levels of glutamine and proline and contain large regions of repetitive sequences. While most gamma gliadins are monomeric proteins containing eight conserved cysteine residues, some contain an additional cysteine residue that enables them to be linked with other gluten proteins into large polymers that are critical for flour quality. The ability to differentiate among the gamma gliadins is important for studies of wheat flour quality because proteins with similar sequences can have different effects on functional properties. The complement of gamma gliadin genes expressed in the wheat cultivar Butte 86 was evaluated by analyzing publicly available expressed sequence tag (EST) data. Eleven contigs were assembled from 153 Butte 86 ESTs. Nine of the contigs encoded full-length proteins and four of the proteins contained nine cysteine residues. Only one of the encoded proteins was a perfect match with a sequence reported in NCBI. Contigs from four different publicly available EST assemblies encoded proteins that were perfect matches with some, but not all, of the Butte 86 gamma gliadins and the complement of identical proteins was different for each assembly. A specialized database that included the sequences of Butte 86 gamma gliadins was constructed for identification of flour proteins by tandem mass spectrometry (MS/MS). In a pilot experiment, proteins corresponding to six Butte 86 gamma gliadin contigs were distinguished by MS/MS, including one containing the extra cysteine residue. Two other proteins were identified as one of two closely related Butte 86 proteins but could not be distinguished unequivocally. Unique peptide tags specific for Butte 86 gamma gliadins are reported. Inclusion of cultivar-specific gamma gliadin sequences in databases maximizes the number and quality of peptide identifications and increases sequence coverage of these gamma gliadins by MS/MS. This approach makes it possible to distinguish closely related proteins, to associate individual proteins with sequences of specific genes, and to evaluate proteomic data in a biological context to better address questions about wheat flour quality.
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影响因子：
3.4
作者：
Mamone, G;Addeo, F;Ferranti, P
通讯作者：
Ferranti, P
影响因子：
3.8
作者：
Pistón, F;Dorado, G;Barro, F
通讯作者：
Barro, F
影响因子：
3.4
作者：
Gobaa, Samy;Bancel, Emmanuelle;Branlard, Gerard
通讯作者：
Branlard, Gerard
影响因子：
3.4
作者：
Muccilli, V;Cunsolo, V;Lafiandra, D
通讯作者：
Lafiandra, D
DOI：
10.1007/s00438-008-0412-8
发表时间：
2009-03
期刊：
Molecular genetics and genomics : MGG
影响因子：
--
作者：
Vaccino P;Becker HA;Brandolini A;Salamini F;Kilian B
通讯作者：
Kilian B