IDENTIFICATION AND DETERMINATION OF COPPER-PROTEIN AND ZINC-PROTEIN COMPLEXES IN BLOOD-PLASMA AFTER CHROMATOGRAPHIC-SEPARATION ON DEAE-SEPHAROSE CL-6B
IDENTIFICATION AND DETERMINATION OF COPPER-PROTEIN AND ZINC-PROTEIN COMPLEXES IN BLOOD-PLASMA AFTER CHROMATOGRAPHIC-SEPARATION ON DEAE-SEPHAROSE CL-6B
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DOI：
10.1039/an9840900871
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发表时间：
1984-01-01
期刊：
影响因子：
4.2
通讯作者：
HODGKINSON, A
中科院分区：
文献类型：
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作者：
CHILVERS, DC;DAWSON, JB;HODGKINSON, A
使用恒定离子强度的缓冲液在 DEAE-Sepharose CL-6B 柱上分离血浆蛋白。通过径向免疫扩散、紫外线吸收和使用纯化的蛋白质“标记”来确定各个组分中蛋白质的身份。通过原子吸收光谱法确定铜和锌的浓度。分级血浆中铜和锌的平均回收率分别为 100.7 ± 4.5 和 100.3 ± 3.8%。同一样品重复分离的变异系数对于铜蓝蛋白-铜为1.3%，对于白蛋白-锌为2.7%，对于α2-巨球蛋白-锌为6.0%。锌主要与白蛋白 (80–90%) 和 α2-巨球蛋白 (10–20%) 结合；其余部分（<3%）与视黄醇结合蛋白复合物相关并且具有较低的相对分子质量分数。在未受污染的血浆中，没有发现锌与 α2-HS-糖蛋白结合的证据。铜主要与铜蓝蛋白 (85-95%) 和白蛋白 (5-15%) 相关。在一些样品中检测到第三个铜峰（<3%），显示出与铜硫因样蛋白质相似的特性。白蛋白-铜复合物比白蛋白-锌复合物洗脱得晚，表明这两种金属通过不同的机制与白蛋白结合。
Plasma proteins were separated on columns of DEAE-Sepharose CL-6B, using a buffer of constant ionic strength. The identities of proteins in individual fractions were determined by radial immunodiffusion, UV absorbance and the use of purified protein ‘markers.’ Copper and zinc concentrations were determined by atomic-absorption spectroscopy. Mean recoveries of copper and zinc in fractionated plasma were 100.7 ± 4.5 and 100.3 ± 3.8%, respectively. The coefficients of variation of replicate separations of the same sample were 1.3% for caeruloplasmin-copper, 2.7% for albumin-zinc and 6.0% for α2-macroglobulin-zinc. Zinc was bound mainly to albumin (80–90%) and α2-macroglobulin (10–20%); the remainder (<3%) was associated with the retinol-binding protein complex and with a low relative molecular mass fraction. No evidence could be found for zinc binding to α2-HS-glycoprotein in uncontaminated plasma. Copper was associated mainly with caeruloplasmin (85–95%) and albumin (5–15%). A third copper peak, (<3%) showing properties similar to a copper-thionein like protein, was detected in some samples. The albumin-copper complex eluted later than the albumin-zinc complex, suggesting that these two metals bind to albumin by different mechanisms.