Shewry P R, Parmar S, Field J M
AFRC Institute of Arable Crops Research, Rothamsted Experimental Station, Harpenden, Herts, UK.
Electrophoresis. 1988 Nov;9(11):727-37. doi: 10.1002/elps.1150091108.
Three complementary two-dimensional systems for the analysis of cereal prolamins are described. These are electrophoresis at pH 3.1 followed by electrophoresis at pH 9.2, isoelectric focusing (IEF) followed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and SDS-PAGE under non-reducing conditions followed by SDS-PAGE with reduction. They together give information on the pIs, Mrs and charge properties of the individual prolamin components, and on their interactions to form oligomers stabilized by inter-chain disulphide bonds. The three systems are then compared for their effectiveness in analysing unreduced prolamin I fractions from wheat and rye, and the IEF/SDS-PAGE system for analysing reduced and pyridylethylated prolamin fractions from all the major cereals. Finally, applications of the systems in biochemical and genetic studies are discussed and illustrated with three examples: analysis of the structural relationships of the S-rich prolamins (B hordeins and gamma-type hordeins) of barley, determination of the interactions of wheat and rye prolamin subunits in a 2RS/2BL translocation line, and the mapping of genes for alpha-type prolamins in the wild grass Haynaldia villosa.
本文描述了三种用于分析谷物醇溶蛋白的互补二维系统。这些系统分别是：先在pH 3.1条件下进行电泳，然后在pH 9.2条件下进行电泳；先进行等电聚焦（IEF），然后进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）；在非还原条件下进行SDS-PAGE，然后在还原条件下进行SDS-PAGE。它们共同提供了关于各个醇溶蛋白组分的等电点、分子量和电荷性质的信息，以及它们通过链间二硫键相互作用形成寡聚体的信息。然后比较了这三种系统在分析小麦和黑麦未还原醇溶蛋白I组分方面的有效性，以及IEF/SDS-PAGE系统在分析所有主要谷物的还原和吡啶基乙基化醇溶蛋白组分方面的有效性。最后，讨论了这些系统在生化和遗传研究中的应用，并通过三个例子进行说明：分析大麦富含S的醇溶蛋白（B醇溶蛋白和γ型醇溶蛋白）的结构关系，确定2RS/2BL易位系中小麦和黑麦醇溶蛋白亚基的相互作用，以及定位野生草本植物绒毛偃麦草中α型醇溶蛋白的基因。
