Das A K, Bhattacharya R, Kundu M, Chakrabarti P, Basu J
Department of Chemistry, Bose Institute, Calcutta, India.
Eur J Biochem. 1994 Sep 1;224(2):575-80. doi: 10.1111/j.1432-1033.1994.00575.x.
Protein 4.2 is a major protein of the human erythrocyte membrane. It has previously been shown to be N-myristoylated. After labeling of intact human erythrocytes with [3H]palmitic acid, radioactivity was found to be associated with protein 4.2 by immunoprecipitation of peripheral membrane proteins extracted at pH 11 from ghosts with anti-(4.2) sera, followed by SDS/PAGE and fluorography. The fatty acid linked to protein 4.2 was identified as palmitic acid after hydrolysis of protein and thin-layer chromatography of the fatty acid extracted in the organic phase. Protein 4.2 could be depalmitoylated with hydroxylamine, suggesting a thioester linkage. Depalmitoylated protein 4.2 showed significantly decreased binding to protein-4.2-depleted membranes, compared to native protein 4.2.
蛋白质4.2是人类红细胞膜的一种主要蛋白质。此前已证明它会发生N-肉豆蔻酰化。在用[3H]棕榈酸标记完整的人类红细胞后,通过用抗(4.2)血清从pH 11的血影中提取外周膜蛋白进行免疫沉淀,随后进行SDS/聚丙烯酰胺凝胶电泳和荧光自显影,发现放射性与蛋白质4.2相关。在蛋白质水解并用有机相提取的脂肪酸进行薄层色谱分析后,与蛋白质4.2相连的脂肪酸被鉴定为棕榈酸。蛋白质4.2可用羟胺去棕榈酰化,提示存在硫酯键。与天然蛋白质4.2相比,去棕榈酰化的蛋白质4.2与耗尽蛋白质4.2的膜的结合显著减少。