Purification and characterization of the acid soluble 26-kDa polypeptide from soybean seeds.
Purification and characterization of the acid soluble 26-kDa polypeptide from soybean seeds.
复制标题
大豆种子中酸溶性 26-kDa 多肽的纯化和表征。
DOI：
10.1271/bbb.61.1286
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
K. Harada
中科院分区：
文献类型：
--
作者：
M. Momma;K. Haraguchi;M. Saito;K. Chikuni;K. Harada
采用sds -聚丙烯酰胺凝胶电泳（SDS-PAGE）对日本品种大豆种子中的乳清蛋白进行了分析。在11个大豆品种中，3个绿色大豆和1个黑色大豆缺少在所有黄色大豆中发现的26 kda条带。本文暂将26-kDa蛋白命名为AS26k（酸溶性26-kDa蛋白）。AS26k蛋白是从Glycine max cv中纯化得到的。通过30-35%饱和硫酸铵分选、S Sepharose Fast Flow离子交换层析、Sephadex G-100凝胶过滤、苯基Sepharose CL-4B疏水层析4个步骤提纯黄豆纳豆。纯化的AS26k用来自金黄色葡萄球菌或CNBr的V8蛋白酶裂解。裂解后的多肽含有DEYGNPV和(M)DKIKEKLPG两个典型的脱氢基序序列，以及与豌豆脱氢基相似的19个氨基酸序列。原生AS26k在凝胶过滤下的分子量为32 kDa，在二维PAGE上的pl为7.2。AS26k与其他脱水剂和胚胎发育晚期丰蛋白（LEA）相似，具有丰富的亲水性氨基酸，热稳定性好。这些结果表明AS26k是大豆种子中II族LEA蛋白的脱氢蛋白。
Whey proteins from soybean seeds of Japanese varieties were analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Among 11 varieties of soybean, three green and one black soybeans lacked a 26-kDa band that was found in all yellow soybeans. In this paper, the 26-kDa protein was named AS26k (acid soluble 26-kDa protein) temporarily. The AS26k protein was purified from Glycine max cv. Nattosyoryu, which is yellow soybean, through four purification steps: 30-35% saturated ammonium sulfate fractionation, ion exchange chromatography on S Sepharose Fast Flow, gel filtration on Sephadex G-100, and hydrophobic chromatography on phenyl Sepharose CL-4B. Purified AS26k was cleaved with V8 proteinase from Staphylococcus aureus or CNBr. The cleaved polypeptide contained two typical dehydrin motif sequences: DEYGNPV and (M)DKIKEKLPG, and a 19 amino acids sequence similar to a pea dehydrin. Native AS26k had a molecular mass of 32 kDa on gel filtration and a pl of 7.2 on two-dimensional PAGE. Similarly to other dehydrins and late embryogenesis abundant (LEA) proteins, AS26k was rich in hydrophilic amino acids, and highly heat stable. These results showed that AS26k was a dehydrin, a group II LEA protein in soybean seeds.