Frenkel E J, Van den Beld B, Van Oost B A, Marx J J
Biochim Biophys Acta. 1983 Jun 15;745(2):202-8. doi: 10.1016/0167-4838(83)90050-x.
Ferritin was purified from rabbit livers either by heat treatment and immunoaffinity chromatography, or by immunoaffinity chromatography alone. The immunoreactivity of ferritin with antibodies raised against heat-treated ferritin was significantly higher for heat-treated preparations than for non-heated preparations. The amount of ferritin protein could be estimated with equal reliability by the assay according to Lowry et al. and by nitrogen determination. Heat treatment favoured the L-subunit-rich ferritin fraction, as measured by densitometric scanning of SDS gradient-pore polyacrylamide gels. Amino acid analysis showed small changes in the amounts of valine, isoleucine and histidine in the heat-treated ferritin, possibly due to selective partial degradation of H-subunit-rich forms of ferritin. These results illustrate that heat treatment, which is a commonly used step in most purification procedures, induces partial denaturation of the ferritin molecules.
铁蛋白可通过热处理和免疫亲和层析法从兔肝脏中纯化得到，也可仅通过免疫亲和层析法纯化。与针对热处理铁蛋白产生的抗体相比，热处理制剂中铁蛋白的免疫反应性明显高于未加热制剂。根据Lowry等人的方法进行的测定以及氮含量测定，均可同等可靠地估计铁蛋白的含量。通过SDS梯度孔聚丙烯酰胺凝胶的光密度扫描测量发现，热处理有利于富含L亚基的铁蛋白组分。氨基酸分析表明，热处理铁蛋白中缬氨酸、异亮氨酸和组氨酸的含量有微小变化，这可能是由于富含H亚基的铁蛋白形式发生了选择性部分降解。这些结果表明，热处理作为大多数纯化程序中常用的步骤，会导致铁蛋白分子发生部分变性。
