Three-dimensional structure of the water-insoluble protein crambin in doidecylphosphocholine micelles and its minimal solvent-exposed surface
Three-dimensional structure of the water-insoluble protein crambin in doidecylphosphocholine micelles and its minimal solvent-exposed surface
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DOI：
10.1021/ja057773d
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发表时间：
2006-04-05
影响因子：
15
通讯作者：
Markley, JL
中科院分区：
文献类型：
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作者：
Ahn, HC;Juranic, N;Markley, JL
我们选择了一种疏水和不溶性蛋白质，最初从植物Crambe abyssinica的种子中分离出来，作为膜相关蛋白的核磁共振研究模型。我们生产了同位素标记的crambin(P22, L25)（含有Pro22和Leu25的crambin变体），作为与葡萄球菌核酸酶的可切割融合，并通过一种已被证明成功地生产具有多个二硫键的蛋白质的方法重新折叠蛋白质。我们使用核磁共振光谱测定了两种模拟膜环境下蛋白质的三维结构：在混合水-有机溶剂（75%/25%，丙酮/水）和在DPC胶束中。将样品置于混合溶剂中，可以测定(>(NHOC)- o -…
We chose crambin, a hydrophobic and water-insoluble protein originally isolated from the seeds of the plant Crambe abyssinica, as a model for NMR investigations of membrane-associated proteins. We produced isotopically labeled crambin(P22, L25) (variant of crambin containing Pro22 and Leu25) as a cleavable fusion with staphylococcal nuclease and refolded the protein by an approach that has proved successful for the production of proteins with multiple disulfide bonds. We used NMR spectroscopy to determine the three-dimensional structure of the protein in two membrane-mimetic environments: in a mixed aqueous-organic solvent (75%/25%, acetone/water) and in DPC micelles. With the sample in the mixed solvent, it was possible to determine (>(NHOC)-O-...