Ligand interaction of human alpha 2-macroglobulin-alpha 2-macroglobulin receptor studied by partitioning in aqueous two-phase systems.
Alpha 2-macroglobulin (alpha 2-M) is a major proteinase inhibitor in human blood and tissue. Besides its antiproteolytic potential, alpha 2-M was found to modulate antigen- and mitogen-driven immune responses and cell growth by binding and transporting distinct cytokines, growth factors and hormones. The inhibitor is cleared from circulation by binding to a multifunctional cellular receptor present on different cell types. Alpha 2-M, as well as its receptor, are capable of binding a variety of ligands. In the present study we have applied aqueous two-phase systems to analyze the interaction of IL-1 beta and alpha 2-M receptor to different forms of alpha 2-M. The partition of IL-1 beta was changed by addition of transformed alpha 2-M to the two-phase systems rather than by the native inhibitor. The interaction between IL-1 beta and alpha 2-M was enhanced by divalent cations. In addition, the complex formation between 125I-labelled receptor and alpha 2-M could clearly be demonstrated by partitioning. In the presence of divalent cations, transformed alpha 2-M, in contrast to the native inhibitor, effectively changed the partition of the receptor. However, the observed alteration of the partition coefficient was found to be less compared with the values obtained by partitioning of the receptor in the presence of whole plasma containing the inhibitor in equivalent concentrations. The results indicate that other components of the plasma exist which competitively bind to the receptor but independent of Ca2+-ions.
Birkenmeier
G
G
Institute of Biochemistry, Medical Faculty, University of Leipzig, Germany.
Kunath
M
M
D000818:Animals; D001798:Blood Proteins / Q000737:chemistry / Q000378:metabolism; D002118:Calcium / Q000737:chemistry; D002413:Cations, Divalent / Q000737:chemistry; D002614:Chelating Agents / Q000737:chemistry; D003124:Colorimetry; D003911:Dextrans / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004492:Edetic Acid / Q000737:chemistry; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D007375:Interleukin-1 / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D007457:Iodine Radioisotopes; D008024:Ligands; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008744:Methylamines / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D011817:Rabbits; D011971:Receptors, Immunologic / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D011973:Receptors, LDL / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D012997:Solvents / Q000737:chemistry; D013501:Surface-Active Agents / Q000737:chemistry; D014867:Water / Q000737:chemistry; D015032:Zinc / Q000737:chemistry; D000511:alpha-Macroglobulins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*
D001798:Blood Proteins; D002413:Cations, Divalent; D002614:Chelating Agents; D003911:Dextrans; D007106:Immune Sera; D007375:Interleukin-1; D007457:Iodine Radioisotopes; D008024:Ligands; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008744:Methylamines; D011971:Receptors, Immunologic; D011973:Receptors, LDL; D012997:Solvents; D013501:Surface-Active Agents; D000511:alpha-Macroglobulins; D014867:Water; D011092:Polyethylene Glycols; D004492:Edetic Acid; C027451:methylamine; D015032:Zinc; D002118:Calcium