Workman E F, Graham G, Bates G W
Biochim Biophys Acta. 1975 Aug 13;399(2):254-64. doi: 10.1016/0304-4165(75)90256-1.
The transfer of iron from transferrin to the developing erythrocyte is a research area of high interest and considerable controversy. We have found that the results of transferrin-reticulocyte incubation studies are quite sensitive to the experimental procedures that are utilized. Reticulocytosis has been induced in rabbits by phelbotomy and phenylhydrazine injections. While the latter gives a higher reticulocyte count, the cells appear to exhibit an altered transferrin-membrane interaction. Transferrin has been iodinated by published methods utilizing chloramine-T and molecular iodine. The iodotransferrin products exhibit the same iron donation ability, however, evidence was found that the chloramine-T treatment leads to a nonspecific binding of transferrin to the reticulocyte. The means of saturating transferrin with 59Fe is also of prime importance. Fe(NH4)2(SO4)2 and especially FeCl3 were found to yield nonspecifically bound iron when added to transferrin or serum. This artifact was reflected in an altered transferrin-reticulocyte interaction. Using what we believe to be optimal conditions, the effect of serum on the transferrin-reticulocyte system was re-examined. The results clearly indicated an enhancement of iron uptake by reticulocytes in the presence of serum, as well as an accelerated incorporation of iron by the cytoplasmic fraction.
铁从转铁蛋白向发育中的红细胞的转移是一个备受关注且存在诸多争议的研究领域。我们发现，转铁蛋白 - 网织红细胞孵育研究的结果对所采用的实验程序非常敏感。通过放血和注射苯肼在兔子身上诱导出了网织红细胞增多症。虽然后者能使网织红细胞计数更高，但这些细胞似乎表现出转铁蛋白 - 膜相互作用的改变。已采用利用氯胺 - T和分子碘的公开方法对转铁蛋白进行碘化。碘化转铁蛋白产物表现出相同的铁捐赠能力，然而，有证据表明氯胺 - T处理会导致转铁蛋白与网织红细胞的非特异性结合。用59Fe使转铁蛋白饱和的方法也至关重要。当将硫酸亚铁铵，特别是氯化铁添加到转铁蛋白或血清中时，会产生非特异性结合的铁。这种假象反映在转铁蛋白 - 网织红细胞相互作用的改变上。使用我们认为是最佳的条件，重新研究了血清对转铁蛋白 - 网织红细胞系统的影响。结果清楚地表明，在有血清存在的情况下，网织红细胞对铁的摄取增强，并且细胞质部分对铁的掺入加速。
