摘要
在TSKgel-Chelate-5PW柱上用固定Cu2^(2+)离子的螯合亲和色谱法(HP-IMAC)研究了Fe(Ⅲ)同去铁转铁蛋白的结合,讨论了所固定的几种金属离子对保留时间和柱寿命的影响;在生理pH条件下分离出了非饱和铁转铁蛋白的4种分子形式:FeNTf,TfFec,FeNTfFec(或Fe2Tf)和Apo-Tf。
he binding of Fe(Ⅱ) by Apo-transferrin has been investigated with high-performance immobilized metal ion affinity chromatography on a Cu (Ⅱ)-loaded TSK gel Chelate-5PW column by elution with linear gradient of imidazole under phymological PH condition.The influences of several loaded metals on the retention time and the column life were discussed. The column life and the affinity of the protein for the immobilized metal ions follow the order Cu ̄(2+) >Ni ̄(2+)>Zn ̄(2+) and the Cu(Ⅱ)-loaded column gives the best resolution. This elution procedure made it possible to use the column over 100 runs when the metal ion Cu ̄(2+) is still immobilized.Under physiological PH condition the transferrin (Tf) which contains iron (not iron-saturated) has been separated into four types of molecules: Fe_NTf, the binding of Fe(Ⅲ) to Tf at the N-terminal. domain I Tffe_c, the binding of Fe(Ⅲ) to Tf at the C-terminal domain f Fe_N TfFe_c (or Fe_2Tf), the binding of Fe(Ⅲ) to Tf on both sites;and Apo-Tf.The foundation of separation method of the four forms of molecules of human transferrin under the physiological PH condition has a great importance to the studies of the binding of the trace ferric iron and other trace metals to human transferrin and their physiological effect.
出处
《色谱》
CAS
CSCD
北大核心
1996年第3期176-178,共3页
Chinese Journal of Chromatography
关键词
螯合亲和色谱
转铁蛋白
铁
色谱分离
immobilized metal ion affinity chromatography, transferrin, apo-transferrin, iron