DETERGENT REMOVAL DURING MEMBRANE RECONSTITUTION
DETERGENT REMOVAL DURING MEMBRANE RECONSTITUTION
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DOI：
10.1016/0005-2736(80)90537-4
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发表时间：
1980-01-01
期刊：
影响因子：
--
通讯作者：
SEGREST, JP
中科院分区：
文献类型：
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作者：
ALLEN, TM;ROMANS, AY;SEGREST, JP
考察了几种不同的膜蛋白重构过程中去除率的影响。研究的重构方法包括乙醇注射-透析、洗涤剂透析和洗涤剂-凝胶过滤。乙醇注射透析法中，.apprx。即使在广泛透析（150小时）后，仍保留70分子乙醇/1000分子磷脂。透析去污剂的效率取决于去污剂。即使对于脱氧胆酸钠，一种具有较大临界胶束浓度的洗涤剂，也有大约0.01%的。7分子脱氧胆酸盐/1000分子磷脂被双分子层保留，即使经过广泛（310小时）透析。用凝胶过滤（Sephadex G-200或G-50）去除脱氧胆酸、胆酸和Triton X-100比透析去除更有效；经过1个色谱柱后，每1000个磷脂分子中只保留10个脱氧胆酸盐分子，只需要几个小时。在Sephadex柱上1次传代去除乙醇的效率低于广泛透析。通过传递或透析bio - beads SM-2去除Triton X-100的结果与通过Sephadex G-200或G-50去除Triton X-100的结果相似。利用凝胶过滤技术。脱氧胆酸钠和一系列磷脂之间对糖蛋白疏水肽T（is）的竞争可能是获得蛋白质-脂质亲和力定量测量的一种手段。T（is）肽与磷脂酰肌醇>磷脂酰胆碱>磷脂酰丝氨酸具有相对的脂质亲和力。
Efficiency of detergent removal during the course of several different procedures for membrane protein reconstitution was examined. Reconstitution methods studied include ethanol injection-dialysis, detergent dialysis and detergent-gel filtration. In the ethanol injection-dialysis method, .apprx. 70 molecules of ethanol/1000 molecules of phospholipid are retained even after extensive (150 h) dialysis. Efficiency of detergent removal by dialysis depends upon the detergent. Even for sodium deoxycholate, a detergent possessing a large critical micelle concentration, there are .apprx. 7 molecules of deoxycholate/1000 molecules of phospholipid retained by the bilayer even after extensive (310 h) dialysis. Detergent removal by gel filtration (Sephadex G-200 or G-50) of deoxycholate, cholic acid and Triton X-100 is more efficient than removal by dialysis; as few as 10 molecules of deoxycholate are retained/1000 molecules of phospholipid after 1 column passage, taking only a few hours. Ethanol was less efficiently removed by 1 passage over a Sephadex column than by extensive dialysis. Removal of Triton X-100 by passage over, or dialysis against, Biobeads SM-2 resulted in a similar level of detergent retention to that found by passage over Sephadex G-200 or G-50. Utilizing gel-filtration techniques. The competition for the hydrophobic peptide of glycophorin, T(is), between sodium deoxycholate and a series of phospholipids as a possible means of obtaining a quantitative measure of protein-lipid affinity. The T(is) peptide has a relative lipid affinity of phosphatidylinositol > phosphatidylcholine > phosphatidylserine.