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首页> 外文期刊>Biological chemistry >Gel Chromatographic Characterization of the Hydrophobic Interaction of Glycosylphosphatidylinositol-Alkaline Phosphatase with Detergents
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Gel Chromatographic Characterization of the Hydrophobic Interaction of Glycosylphosphatidylinositol-Alkaline Phosphatase with Detergents

机译：糖基磷脂酰肌醇-碱性磷酸酶与洗涤剂的疏水相互作用的凝胶色谱表征

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摘要

The interaction of a glycosylphosphatidylinositol (GPI) protein with different detergents was studied for the first time with a purified protein. Four differently hydrophobic fractions of GPI-alkaline phosphatase (GPI-AP) from calf intestine were used as model proteins. The mode of interaction was determined by investigating (i) the self-aggregation behaviour of the GPI-AP fractions, (ii) the interference of detergents with GPI-AP binding to octyl-Sepharose, and (iii) the elution of GPI-AP bound to octyl-Sepharose. It was shown that polyoxyethylene-type detergents surprisingly interact much stronger than n-octylglucoside with GPI-AP, which is in contrast to the known behaviour of GPI-proteins in natural membranes. Gel filtration chromatography of Triton X-100 at concentrations above the critical micellar concentration yields three different micelle species with apparent molecular weights of about 166, 54, and 16 kDa. GPI-AP fraction II, which is shown to bear only one anchor per dimer, does not bind to any of these micelles. We demonstrate that a complex is formed containing about 150 Triton X-100 molecules and about 4700 molecules of water per molecule of GPI-AP dimer. The experimental findings are in accordance with a simple geometrical model based on the physical data of fatty acids and the arrangement, mean size, and shape of Triton X-100 molecules.
机译：糖基磷脂酰肌醇（GPI）蛋白与不同去污剂的相互作用首次与纯化蛋白进行了研究。来自小牛肠的GPI-碱性磷酸酶（GPI-AP）的四个不同疏水级分用作模型蛋白。通过研究（i）GPI-AP组分的自聚集行为，（ii）去污剂对GPI-AP与辛基-琼脂糖结合的干扰以及（iii）GPI-AP的洗脱来确定相互作用的方式与辛基-琼脂糖结合。结果表明，聚氧乙烯型去污剂与GPI-AP的相互作用比正辛基葡糖苷强得多，这与天然膜中GPI蛋白的已知行为相反。在高于临界胶束浓度的浓度下，Triton X-100的凝胶过滤色谱法产生三种不同的胶束物质，其表观分子量约为166、54和16 kDa。 GPI-AP级分II（每个二聚体仅带有一个锚）不与任何这些胶束结合。我们证明形成的复合物每个分子的GPI-AP二聚体包含约150个Triton X-100分子和约4700个水分子。实验结果与基于脂肪酸物理数据以及Triton X-100分子的排列，平均大小和形状的简单几何模型一致。

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