Hoefkens P, de Smit M H, de Jeu-Jaspars N M, Huijskes-Heins M I, de Jong G, van Eijk H G
Department of Chemical Pathology, Erasmus University, Rotterdam, The Netherlands.
Int J Biochem Cell Biol. 1996 Sep;28(9):975-82. doi: 10.1016/1357-2725(96)00057-x.
Recombinant human transferrin as well as N- and C-terminal half-transferrins, produced in Escherichia coli, are deposited in inclusion bodies by the bacteria. The isolation and purification of the recombinant proteins from these inclusion bodies are described here. The amino acid compositions and N-terminal sequences of the proteins were determined, and found to be in agreement with the known protein structure of human serum transferrin. Renaturation of the recombinant proteins is described, resulting in water-soluble iron-binding molecules. Iron binding was confirmed by 59Fe labelling, absorption spectrophotometry and EPR spectrometry.
在大肠杆菌中产生的重组人转铁蛋白以及N端和C端半转铁蛋白会被细菌沉积在包涵体中。本文描述了从这些包涵体中分离和纯化重组蛋白的方法。测定了这些蛋白的氨基酸组成和N端序列，发现与已知的人血清转铁蛋白的蛋白质结构一致。文中还描述了重组蛋白的复性过程，最终得到了水溶性的铁结合分子。通过59Fe标记、吸收分光光度法和电子顺磁共振光谱法证实了铁结合。
