Robinson J B, Strottmann J M, Wick D G, Stellwagen E
Proc Natl Acad Sci U S A. 1980 Oct;77(10):5847-51. doi: 10.1073/pnas.77.10.5847.
Anionic dye affinity chromatography is commonly unproductive in the presence of nonionic detergents used to extract particulate proteins. Using lactate dehydrogenase as a model protein, Cibacron blue F3GA as a model dye, and Triton X-100 as a model detergent, we find that the dye is encapsulated in nonionic detergent micelles, rendering the dye incapable of ligation with the enzyme. However, the dye can be liberatd from the micelles without altering the nonionic detergent concentration by addition of an anionic detergent, such as deoxycholate or sodium dodecyl sulfate, forming mixed anionic/nonionic micelles that displace the anionic dye. Encapsulation of the anionic detergents prevents their activity as protein denaturants. These observations have been successfuly translated to the dye affinity chromatography of a detergent extract of brain particulate cyclic nucleotide phosphodiesterase.
在用于提取颗粒蛋白的非离子去污剂存在的情况下，阴离子染料亲和色谱通常效率不高。以乳酸脱氢酶作为模型蛋白、汽巴克隆蓝F3GA作为模型染料、Triton X - 100作为模型去污剂，我们发现该染料被包裹在非离子去污剂胶束中，致使染料无法与酶结合。然而，通过添加阴离子去污剂，如脱氧胆酸盐或十二烷基硫酸钠，形成混合阴离子/非离子胶束来取代阴离子染料，可在不改变非离子去污剂浓度的情况下从胶束中释放出染料。阴离子去污剂的包裹可防止其作为蛋白质变性剂的活性。这些观察结果已成功应用于脑颗粒环核苷酸磷酸二酯酶去污剂提取物的染料亲和色谱分析。
