Species specificity of transferein binding,endocytosis and iron internalization by cultured chick myogenic cells

Summary The ability of unlabelled heterologous transferrin to interact with transferrin receptors on developing chick myogenic cells was investigated by measuring their capacity to inhibit the surfacebinding and internalization of¹²⁵I-and⁵⁹Fe-labelled ovotransferrin. Transferrins from rat, rabbit, human, and a species of kangaroo (Macropus fuliginosus) were unable to inhibit either surfacebinding or internalization of labelled ovotransferrin even at concentrations ten times the molar concentration of the ovotransferrin. Transferrins isolated from the serum of a toad (Bufo marinus) and a lizard (Teliqua rugosa), when added at high concentrations, were found to reduce surface-binding of¹²⁵I-Tf by 20–25% but did not inhibit internalization of either¹²⁵I-Tf or⁵⁹Fe. This suggests that the effects of toad and lizard transferrins are due to non-specific binding to the myogenic cells. In contrast, inhibition of both surface-binding and internalization of labelled ovotransferrin was found when myogenic cells were incubated in the presence of the homologous transferrin (ovotransferrin). The species-specificity of transferrin binding, endocytosis and iron internalization did not vary with the state of proliferation or differentiation of the myogenic cells. However, the intracellular iron utilization was found to differ between differentiating presumptive and terminally differentiated myotubes. Internalized⁵⁹Fe was fractioned by gel filtration. In dividing and non-dividing presumptive myoblasts⁵⁹Fe was found to elute in three peaks, two with elution volumes corresponding to ferritin and transferrin and one at greater elution volume than that of myoglobin. In myotubes the same fractions occurred, and in addition some⁵⁹Fe was eluted at the same volume as myoglobin.Abbreviations Tf-Fe ₂ differic transferrin - BSA bovine serum albumin - BSS balanced salt solution - MEM Eagle's modified minimum essential medium - MW molecular weight - BUdR bromodeoxyuridine - Ara-C cytosine arabinoside