Baker B L, Hultquist D E
J Biol Chem. 1978 Feb 25;253(4):1195-200.
A copper.protein complex present in the serum of a hypercupremic myeloma patient has been purified to homogeneity using gel filtration, DEAE-cellulose chromatography, and concanavalin A/Sepharose affinity chromatography. Immunoelectrophoresis and hemagglutination inhibition tests showed the copper-bound protein to be an IgG1-type immunoglobulin with lambda light chains. The immunoglobulin is of normal molecular weight (150,000) with normal size light and heavy chains (28,000 and 56,000, respectively). The carbohydrate portion of the molecule appears to be abnormal in that it interacts with concanavalin A, whereas most immunoglobulins of the gammaG-type do not. The copper in the native copper.IgG complex is in an EPR-indeterminable valence state. Copper was efficiently removed from the copper.IgG complex by dialysis against 0.1 M potassium cyanide. The apo-IgG was separated from the copper.cyanide complex by gel filtration. The copper complex was reconstituted by equilibrating the apo-IgG with 7.7 muM cupric ions.
使用凝胶过滤、DEAE-纤维素色谱法和伴刀豆球蛋白A/琼脂糖亲和色谱法，已将一名高铜血症骨髓瘤患者血清中存在的一种铜-蛋白质复合物纯化至同质状态。免疫电泳和血凝抑制试验表明，与铜结合的蛋白质是一种带有λ轻链的IgG1型免疫球蛋白。该免疫球蛋白具有正常的分子量（150,000）以及正常大小的轻链和重链（分别为28,000和56,000）。该分子的碳水化合物部分似乎异常，因为它能与伴刀豆球蛋白A相互作用，而大多数γG型免疫球蛋白则不能。天然铜-IgG复合物中的铜处于电子顺磁共振无法确定的价态。通过用0.1 M氰化钾透析，铜从铜-IgG复合物中被有效去除。脱辅基IgG通过凝胶过滤与铜-氰化物复合物分离。通过使脱辅基IgG与7.7 μM铜离子平衡来重建铜复合物。
