[Rapid analysis of transferrin-iron complexes by isoelectric focusing and radioactive densitometry on gel]. 1985

J M Legendre, and M P Moineau, and J F Menez, and A Turzo

The analysis of the molecular forms of transferrin-iron complexes with 59Fe, was carried out using an isoelectric focusing method on commercially available polyacrylamide plates (PAG plates pH = 4-6,5, LKB). Depending upon the saturation degree of proteins with iron, apotransferrin (pI = 6,1), transferrin-Fe3+ (pI = 5,8), transferrin-2 Fe3+ (a) (pI = 5,5) and transferrin-2 Fe3+ (b) (pI = 5,3) were put forward. When the protein was saturated with iron, a double band was observed which was assigned to the microheterogeneity of the protein and depends upon the sialic content. The quantitative analysis was carried out by studying 59Fe radioactivity distribution directly on the plate. As a control, a specific staining of proteins was achieved. The interpretation of the results is difficult if such a method is used, as some bands are not attributed. Through an unspecific staining, the UV-visible densitometry is not satisfactory enough. The proposed method allows a fast analysis of transferrin-metal complexes labeled with radioactive metals. Such a method would be adaptable to the study of other complexes as transferrin-67Ga, transferrin 111In in order to determine the binding site of metal on the protein and its metal content.

UI MeSH Term Description Entries
D007501 Iron A metallic element with atomic symbol Fe, atomic number 26, and atomic weight 55.85. It is an essential constituent of HEMOGLOBINS; CYTOCHROMES; and IRON-BINDING PROTEINS. It plays a role in cellular redox reactions and in the transport of OXYGEN. Iron-56,Iron 56
D007504 Iron Radioisotopes Unstable isotopes of iron that decay or disintegrate emitting radiation. Fe atoms with atomic weights 52, 53, 55, and 59-61 are radioactive iron isotopes. Radioisotopes, Iron
D007525 Isoelectric Focusing Electrophoresis in which a pH gradient is established in a gel medium and proteins migrate until they reach the site (or focus) at which the pH is equal to their isoelectric point. Electrofocusing,Focusing, Isoelectric
D011485 Protein Binding The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments. Plasma Protein Binding Capacity,Binding, Protein
D003720 Densitometry The measurement of the density of a material by measuring the amount of light or radiation passing through (or absorbed by) the material. Densitometries
D014168 Transferrin An iron-binding beta1-globulin that is synthesized in the LIVER and secreted into the blood. It plays a central role in the transport of IRON throughout the circulation. A variety of transferrin isoforms exist in humans, including some that are considered markers for specific disease states. Siderophilin,Isotransferrin,Monoferric Transferrins,Serotransferrin,Transferrin B,Transferrin C,beta 2-Transferrin,beta-1 Metal-Binding Globulin,tau-Transferrin,Globulin, beta-1 Metal-Binding,Metal-Binding Globulin, beta-1,Transferrins, Monoferric,beta 1 Metal Binding Globulin,beta 2 Transferrin,tau Transferrin
D014462 Ultrafiltration The separation of particles from a suspension by passage through a filter with very fine pores. In ultrafiltration the separation is accomplished by convective transport; in DIALYSIS separation relies instead upon differential diffusion. Ultrafiltration occurs naturally and is a laboratory procedure. Artificial ultrafiltration of the blood is referred to as HEMOFILTRATION or HEMODIAFILTRATION (if combined with HEMODIALYSIS).

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