title: Binding of alpha 2-macroglobulin-thrombin complexes and methylamine-treated alpha 2-macroglobulin to human blood monocytes.
abstract: The binding of alpha 2-macroglobulin (alpha 2M) to human peripheral blood monocytes was investigated. Monocytes, the precursors of tissue macrophages, were isolated from fresh blood by centrifugal elutriation or density gradient centrifugation. Binding studies were performed using 125I-labeled alpha 2M. Cells and bound ligand were separated from free ligand by rapid vacuum filtration. Nonlinear least-squares analysis of data obtained in direct binding studies at 0 degrees C showed that monocytes bound the alpha 2M-thrombin complex with a Kd of 3.0 +/- 0.9 nM and the monocyte had 1545 +/- 153 sites/cell. Thrombin alone did not compete for the site. Binding was divalent cation dependent. Direct binding studies also demonstrated that monocytes bound methylamine-treated alpha 2M in a manner similar to alpha 2M-thrombin. Competitive binding studies showed that alpha 2M-thrombin and methylamine-treated alpha 2M bound to the same sites on the monocyte. In contrast, native alpha 2M did not compete with alpha 2M-thrombin for the site. Studies done at 37 degrees C suggested that after binding, the monocyte internalized and degraded alpha 2M-thrombin and excreted the degradation products. Receptor turnover and degradation of alpha 2M-thrombin complexes were blocked in monocytes treated with chloroquine, an inhibitor of lysosomal function. Our results indicate that human monocytes have a divalent cation dependent, high-affinity binding site for alpha 2M-thrombin and methylamine-treated alpha 2M which may function to clear alpha 2M-proteinase complexes from the circulation.
authors: Center for Thrombosis and Hemostasis, University of North Carolina, Chapel Hill 27514.
authors: D L
authors: DL
authors: Straight
authors: L
authors: L
authors: Jakoi
authors: P A
authors: PA
authors: McKee
authors: R
authors: R
authors: Snyderman
mesh_terms: D001667:Binding, Competitive; D006801:Humans; D007700:Kinetics; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008744:Methylamines / Q000494:pharmacology*; D009000:Monocytes / Q000187:drug effects / Q000276:immunology*; D011971:Receptors, Immunologic / Q000378:metabolism*; D013917:Thrombin / Q000378:metabolism*; D000511:alpha-Macroglobulins / Q000378:metabolism*
chemical_list: D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008744:Methylamines; D011971:Receptors, Immunologic; D000511:alpha-Macroglobulins; C027451:methylamine; D013917:Thrombin