Bhushan R, Reddy K R
Dept. of Chemistry, University of Roorkee, India.
Int J Pept Protein Res. 1990 May;35(5):406-15. doi: 10.1111/j.1399-3011.1990.tb00066.x.
A subunit of molecular weight 21,000 from arachin, the major peanut protein, was isolated in pure form and primary structure was determined. The subunit was fragmented with CNBr, trypsin, and NBS; the fragments were separated and isolated by PAGE, gel filtration, Dowex treatment, and paper electrophoresis, and Edman degradation on each fragment, including the intact subunit, was performed. The PTH-amino acids thus obtained were identified by UV spectroscopy and TLC. The complete sequence of 176 residues was established by overlapping technique.
从花生主要蛋白质花生球蛋白中分离出了一种分子量为21,000的亚基,并确定了其一级结构。该亚基用溴化氰、胰蛋白酶和N-溴代琥珀酰亚胺进行裂解;裂解片段通过聚丙烯酰胺凝胶电泳(PAGE)、凝胶过滤、Dowex处理和纸电泳进行分离和纯化,对每个片段(包括完整的亚基)进行了埃德曼降解。通过紫外光谱和薄层色谱法对由此获得的苯乙内酰硫脲氨基酸进行了鉴定。通过重叠技术确定了176个残基的完整序列。