Appleby C A, Nicola N A, Hurrell J G, Leach S J
Biochemistry. 1975 Oct 7;14(20):4444-50. doi: 10.1021/bi00691a016.
An improved separation procedure is described for isolating five leghemoglobin components from the nodules of soybean plants. After a preliminary oxidation with ferricyanide, and separation from endogenous nicotinate at pH 9.2, the ferrileghemoglobins are separated by DEAE-cellulose chromatography using gradient elution with acetate buffer (pH 5.2). The components have been characterized by their acetate and nicotinate binding affinities, gel electrophoretic, visible, and circular dichroic spectra in the ultraviolet, Soret and visible regions. Two formerly unresolved components of leghemoglobin c have indistinguishable circular dichroic, electrophoretic, and ligand binding properties, but differ in their spin states as judged by their visible spectra, their amino acid analyses, and their tryptic maps.
本文描述了一种改进的分离方法，用于从大豆植株的根瘤中分离出五种豆血红蛋白成分。在用铁氰化物进行初步氧化，并在pH 9.2条件下与内源性烟酸分离后，使用乙酸盐缓冲液（pH 5.2）梯度洗脱，通过DEAE-纤维素色谱法分离高铁豆血红蛋白。这些成分已通过它们与乙酸盐和烟酸的结合亲和力、凝胶电泳、紫外、索雷特和可见光区域的可见光谱以及圆二色光谱进行了表征。豆血红蛋白c的两个以前未分离的成分具有难以区分的圆二色性、电泳和配体结合特性，但根据它们的可见光谱、氨基酸分析和胰蛋白酶图谱判断，它们的自旋状态不同。
