2-DE技术中疏水性和碱性蛋白质的研究进展

双向凝胶电泳(2-DE)具有高分辨率、高通量等特点，已被广泛地用于蛋白质组的分离．但是它在分离疏水性蛋白质和碱性蛋白质时却遇到了极大的挑战．然而，疏水性与碱性蛋白质在全蛋白质中占相当大的比例，且具有很重要的生物学意义．因而，近年来，越来越多的研究者将目标瞄准这些蛋白质，并且取得了一些令人鼓舞的进展：用亚细胞预分离技术，顺序提取法等方法来富集疏水性蛋白质，用一些新的有效的增溶剂如硫脲，ASB一14等来改善疏水性蛋白质的溶解，应用这些技术2一DE可分辨出总平均疏水值达O．80的蛋白质；在碱性蛋白质分离方面，通过等电聚焦预处理，使用窄pH梯度胶条等大大地改善了碱性蛋白质在2-DE中的分离，能分辨出等电点达11．7的蛋白质．现对2-DE技术中疏水性和碱性蛋白质分离的研究进展进行综述．

The Application of 2-DE Technique to Hydrophobic Proteins and Alkaline Proteins

Two dimensional gel electrophoresis(2 DE) has the property of high resolution, high throughput, so it has been used for the isolation of proteins widely,but it has continued to be a great challenge to separate hydrophobic proteins and alkaline proteins. However, hydrophobic proteins and alkaline proteins account for big scale in global proteins and they are meaningful in biology. Therefore, recently, more and more researchers have aimed at these proteins, and make some invigorating progress: enriching hydrophobic proteins by subcellular prefractionation, sequential extraction. and using chaotropes such as thiourea, ASB 14 to improve the solubility of hydrophobic proteins. Thus, the proteins with high grand average hydrophobicity (GRAVY) values even up to 0.80 can be detectable now. For improving the separation of alkaline proteins, it is helpful to use isoelectric focusing prefractionation technology, and narrow range immobilized pH gradients strips. Thus, the proteins with high pI even up to 11.7 are detectable now. The recent evolution of the separation of hydrophobic proteins and alkaline proteins by 2 DE technology are reviewed.