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[节节麦种子醇溶蛋白的分离与鉴定]

[Isolation and characterization of gliadins from Aegilops squarrosa seeds].

作者信息

Odintsova T I, Egorov Ts A

出版信息

Biokhimiia. 1989 Mar;54(3):396-408.

PMID:2752062
Abstract

The major gliadin components were isolated from the seeds of the diploid species Aegilops squarrosa, a putative source of polyploid wheat D-genome. The isolation procedure included gel-filtration and reversed-phase high-performance liquid chromatography (HPLC). The purified proteins were characterized by electrophoretic mobility in polyacrylamide gel using acid Al-lactate system and a system containing sodium dodecyl sulfate. The amino acid composition of isolated omega-gliadins was determined. Using covalent chromatography on thiopropyl-Sepharose 6B it was found that omega-gliadins of A. squarrosa contain no SH-groups and/or S-S-bonds. The N-terminal amino acid sequences of A. squarrosa gliadins were determined. omega-Gliadins were found to contain three types of N-terminal amino acid sequences, one of which, SRQ, in hexaploid wheat is encoded by 1B chromosome. It was shown that some omega-gliadins of A. squarrosa have blocked N-terminal amino acids. The major component of the gamma-fraction was found to contain an N-terminal sequence of gamma 2 type encoded in polyploid wheat by 1D chromosome. Gliadins with electrophoretic mobility in the beta-zone of the spectrum possess the N-terminal sequence of alpha-type. The results obtained are discussed in terms of the origin of polyploid wheat genomes.

摘要

主要麦醇溶蛋白组分是从二倍体物种节节麦的种子中分离得到的,节节麦被认为是多倍体小麦D基因组的一个来源。分离过程包括凝胶过滤和反相高效液相色谱(HPLC)。通过在聚丙烯酰胺凝胶中使用酸性α-乳酸系统和含有十二烷基硫酸钠的系统进行电泳迁移率分析,对纯化的蛋白质进行了表征。测定了分离得到的ω-麦醇溶蛋白的氨基酸组成。通过在硫丙基-琼脂糖6B上进行共价色谱分析发现,节节麦的ω-麦醇溶蛋白不含SH基团和/或二硫键。测定了节节麦麦醇溶蛋白的N端氨基酸序列。发现ω-麦醇溶蛋白含有三种类型的N端氨基酸序列,其中一种,SRQ,在六倍体小麦中由1B染色体编码。结果表明,节节麦的一些ω-麦醇溶蛋白具有封闭的N端氨基酸。发现γ-组分的主要成分含有由多倍体小麦中的1D染色体编码的γ2型N端序列。在光谱β区具有电泳迁移率的麦醇溶蛋白具有α型N端序列。根据多倍体小麦基因组的起源对所得结果进行了讨论。

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