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Comparative studies of the binding and growth-supportive ability of mammalian transferrins in human cells.
Abstract: The ability of human-derived cells in culture to bind, remove iron from, and grow in the presence of transferrins (Tf) isolated from the sera of species commonly included in tissue culture medium was investigated. Kinetic studies on HeLa cells reveal apparent first-order association rate constants of 0.43 min-1 for human Tf and 0.15 min-1 for equine Tf. Labeled chicken ovo-Tf and fetal bovine Tf were not recognized by the HeLa cells. Competition experiments with HeLa cells that use either isolated Tf or parent serum confirm these findings. Equilibrium binding experiments performed on HeLa cells at 37 degrees C in the presence of 2,4-dinitrophenol to prevent iron removal indicate 1 X 10(6) Tf bound/cell with a dissociation constant (K'D) of 28 nM for human Tf and 182 nM for equine Tf. Equilibrium binding performed at 0 degrees C to prevent endocytosis reveals 4.1-6.7 X 10(5) Tf binding sites/cell with a K'D of 8.3 nM for human Tf and 41.5 nM for equine Tf. Parallel experiments in normal human diploid fibroblast-like MRC-5 cells indicate expression of 0.82-2.78 X 10(5) Tf binding sites/cell with a K'D of 8.2 nM for human and 39.1 nM for equine Tf. Thus, the results of equilibrium binding studies of a more differentiated cell type are consistent with those found for HeLa cells. Fetal bovine Tf was found to compete weakly with labeled human Tf for human receptor on HeLa cells in a soluble receptor assay, with an approximately 500-fold excess needed to reduce binding to half maximal. Iron uptake experiments show an iron donating hierarchy where human greater than horse greater than calf, suggesting that the rate of iron uptake depends on the affinity of receptor for transferrin. Growth experiments involving HeLa cells in chemically defined serum-free medium demonstrate that bovine Tf will support growth as well as human Tf, but at concentrations much higher than are required of human Tf.
Publication Date: 1986-08-01 PubMed ID: 3015988DOI: 10.1002/jcp.1041280217Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Comparative Study
- Journal Article
- Research Support
- U.S. Gov't
- P.H.S.
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This research investigates the binding and growth-supportive qualities of transferrins (a type of protein that carries iron) from different species on human cells. The study found that human cells recognize and better utilize human and horse transferrins, but do not recognize chicken or bovine transferrins. The efficiency of iron uptake by the cells depends on the source of transferrin, with human-transferrin being the most efficient.
Experiments and Findings
- The study began with kinetic investigations on HeLa cells (a strain of human cells commonly used in scientific research), focusing on the binding speed (association rate constants) of human and equine (horse) transferrins. The human transferrin had a faster binding rate.
- Further tests showed that HeLa cells could not recognize chicken or bovine (cow) transferrins – proteins labelled with these transferrins were not detected by the cells.
- When the researchers inhibited the process of iron removal in the HeLa cells and studied the equilibrium binding, they found large numbers of transferrin binding sites. Human transferrin showed higher affinity (ability to bind) to the cell receptors compared to equine transferrin.
- This pattern of results was repeated when the process of endocytosis (cellular intake of substances) was prevented in the cells.
Comparison with Different Cell Types
- Tests on another type of human cell, known as MRC-5 cells, yielded similar results. It confirmed the expression of transferrin binding sites and showed similar relative affinities between human and equine transferrins. This demonstrates that these results are not limited to only HeLa cells but are likely to be more generally applicable.
Competitive Experiments
- Fetal bovine transferrin was found to weakly compete with human transferrin for the binding sites on the HeLa cells. It did so only with a 500-fold excess compared to the human transferrin.
Iron Uptake and Cell Growth
- The study also examined iron uptake by cells depending on the transferrin source. It found a hierarchy with human transferrin resulting in the highest iron intake by the cells, followed by horse and then calf transferrin.
- In growth experiments, bovine transferrin was able to support the growth of HeLa cells similarly to human transferrin, but only at much higher concentrations.
Implications
- The findings have implications for laboratory cell culture methods. It’s necessary to consider the source of transferrin when cultivating human cells in the lab, as some types of transferrin may not be recognized by the cells or may require much higher concentrations to support cell growth.
Cite This Article
APA
Penhallow RC, Brown-Mason A, Woodworth RC.
(1986).
Comparative studies of the binding and growth-supportive ability of mammalian transferrins in human cells.
J Cell Physiol, 128(2), 251-260.
https://doi.org/10.1002/jcp.1041280217 Publication
Researcher Affiliations
MeSH Terms
- Animals
- Binding, Competitive
- Cattle / blood
- Cell Division
- Cell Line
- HeLa Cells
- Horses / blood
- Humans
- Iron / metabolism
- Radioimmunoassay
- Receptors, Cell Surface / metabolism
- Receptors, Transferrin
- Species Specificity
- Transferrin / metabolism
- Transferrin / pharmacology
Grant Funding
- AM21739 / NIADDK NIH HHS
- HL30373 / NHLBI NIH HHS
Citations
This article has been cited 13 times.- Marta CM, Adrian M, Jorge FD, Francisco AM, De Miguel MP. Improvement of an Effective Protocol for Directed Differentiation of Human Adipose Tissue-Derived Adult Mesenchymal Stem Cells to Corneal Endothelial Cells. Int J Mol Sci 2021 Nov 5;22(21).
- Zhang D, Lee HF, Pettit SC, Zaro JL, Huang N, Shen WC. Characterization of transferrin receptor-mediated endocytosis and cellular iron delivery of recombinant human serum transferrin from rice (Oryza sativa L.). BMC Biotechnol 2012 Nov 30;12:92.
- Luck AN, Mason AB. Structure and dynamics of drug carriers and their interaction with cellular receptors: focus on serum transferrin. Adv Drug Deliv Rev 2013 Jul;65(8):1012-9.
- Steere AN, Byrne SL, Chasteen ND, Smith VC, MacGillivray RT, Mason AB. Evidence that His349 acts as a pH-inducible switch to accelerate receptor-mediated iron release from the C-lobe of human transferrin. J Biol Inorg Chem 2010 Nov;15(8):1341-52.
- Wally J, Halbrooks PJ, Vonrhein C, Rould MA, Everse SJ, Mason AB, Buchanan SK. The crystal structure of iron-free human serum transferrin provides insight into inter-lobe communication and receptor binding. J Biol Chem 2006 Aug 25;281(34):24934-44.
- Feder JN. The hereditary hemochromatosis gene (HFE): a MHC class I-like gene that functions in the regulation of iron homeostasis. Immunol Res 1999;20(2):175-85.
- Mason AB, Tam BM, Woodworth RC, Oliver RW, Green BN, Lin LN, Brandts JF, Savage KJ, Lineback JA, MacGillivray RT. Receptor recognition sites reside in both lobes of human serum transferrin. Biochem J 1997 Aug 15;326 ( Pt 1)(Pt 1):77-85.
- Mason AB, Brown SA. Differential effect of iodination of ovotransferrin and its two half-molecule domains on binding to transferrin receptors on chick embryo red blood cells. Biochem J 1987 Oct 15;247(2):417-25.
- Andreesen R, Sephton RG, Gadd S, Atkins RC, De Abrew S. Human macrophage maturation in vitro: expression of functional transferrin binding sites of high affinity. Blut 1988 Aug;57(2):77-83.
- Sorokin LM, Morgan EH. Species specificity of transferrin binding, endocytosis and iron internalization by cultured chick myogenic cells. J Comp Physiol B 1988;158(5):559-66.
- Weller M, Wiedemann P, Moter H, Heimann K. Transferrin and transferrin receptor expression in intraocular proliferative disease. APAAP-immunolabeling of retinal membranes and ELISA for vitreal transferrin. Graefes Arch Clin Exp Ophthalmol 1989;227(3):281-6.
- Weber T, Malakpour-Permlid A, Chary A, D'Alessandro V, Haut L, Seufert S, Wenzel EV, Hickman J, Bieback K, Wiest J, Dirks WG, Coecke S, Oredsson S. Fetal bovine serum: how to leave it behind in the pursuit of more reliable science. Front Toxicol 2025;7:1612903.
- Fraser R, Campbell K, Pokorski P, MacKinnon E, McAllister K, Neves KB, Murphy F. Humanising nanotoxicology: replacement of animal-derived products in the application of integrated approaches to testing and assessment of nanomaterial inhalation hazard. Front Bioeng Biotechnol 2025;13:1526808.
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