Isolation and characterization of beta-glucan receptors on human mononuclear phagocytes.
beta-glucan receptors, with ligand specificity for yeast and fungal carbohydrate polymers, have been studied as phagocytic receptors of human monocytes. To characterize their structure, binding studies were carried out with human U937 cells and a rabbit IgG anti-Id that recognizes epitopes on monocyte beta-glucan receptors. Unstimulated U937 cells specifically bound large amounts of the anti-Id, but almost none of the control anti-isotype. At saturation, the number of anti-Id molecules bound per U937 cell was 2.6 x 10(6) with an apparent Ka of 1.9 x 10(7) M-1. Immunoprecipitates from detergent lysates of surface-radioiodinated U937 cells contained only two membrane proteins with antigenic specificity for the anti-Id, one having a mol wt of 180 kD and the other 160 kD. Both proteins were disulfide-linked and presented, after reduction, as five polypeptides of 95, 88, 60, 27, and 20 kD. Detergent lysates of unlabeled U937 cells, purified by affinity chromatography on anti-Id-Sepharose, yielded the same two nonreduced proteins and five reduction products in slab gels stained with Coomassie blue. In Western blots probed with the anti-Id, the most immunoreactive nonreduced and reduced affinity-purified products were the 160 and 20 kD molecules, respectively. Immunoblots of two-dimensional gels showed the 180 and 160 kD proteins to express a common epitope through disulfide linkage to the 20 kD polypeptide. By immunoblot analysis, U937 cell glucan-binding proteins from detergent lysates contained two cell proteins antigenic for the anti-Id that were indistinguishable from affinity-purified molecules in size and subunit composition. Studies of affinity-purified proteins from detergent lysed human monocytes were characterized by immunoblot analysis and found to be identical to U937 cell beta-glucan receptors. They consisted of two disulfide-linked proteins, with mol wt of 180 and 160 kD, and had in common a 20 kD polypeptide with the anti-Id epitope.
Czop
J K
JK
Department of Medicine, Harvard Medical School, Boston, Massachusetts 02115.
Kay
J
J
D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D002460:Cell Line; D002846:Chromatography, Affinity; D015180:Electrophoresis, Gel, Two-Dimensional; D005936:Glucans / Q000378:metabolism; D006801:Humans; D008970:Molecular Weight; D009000:Monocytes / Q000737:chemistry*; D011233:Precipitin Tests; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000302:isolation & purification*; D011971:Receptors, Immunologic
D000911:Antibodies, Monoclonal; D005936:Glucans; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic; C045268:beta-glucan receptor