title: Phospholipase A2 activity of Fc gamma 2b receptors of thioglycollate-elicited murine peritoneal macrophages.
abstract: The detergent lysate of plastic adherent cell population of thioglycollate-elicited peritoneal exudate cells from 100 individual Swiss mice was subjected to affinity chromatography on two different media, Sepharose coupled to heat-aggregated human IgG (IgG-Sepharose) and Sepharose coupled to the phosphatidylcholine analog, rac-1-(9-carboxyl)nonyl-2-hexadecylglycero-3 -phosphorylcholine (PC-Sepharose). Both IgG- and PC-binding proteins were further purified by Sephadex G-100 gel filtration and isoelectric focusing in the presence of 6M urea. Both IgG- and PC-binding proteins thus purified appear to be homogeneous in size as well as charge properties. The IgG-binding proteins of a molecular weight of 25,000 had an isoelectric point of 4.8, whereas the PC-binding proteins of a molecular weight of 42,000 were more basic and had an isoelectric point of 6.0. Both materials retained their IgG-binding capabilities as judged by their inhibitory capacity of murine EA gamma rosetting systems. The subclass specificities of the IgG- and the PC-binding proteins were for IgG2a and IgG2b, respectively. The PC-binding proteins possessed a typical phospholipase A2 activity, which was maximal at pH 9.5, depended on Ca++, and was specific for cleavage of fatty acid from the sn-2 position of phosphatidylcholine. The binding of aggregated IgG2b to the PC-binding proteins caused the ninefold increase in noted enzymatic activity in the presence of, but not in the absence of, Ca++ (5mM). The IgG-binding proteins on the other hand, lacked any detectable phospholipase A2 activity. Thus, the biochemical and biological properties of the PC- and IgG-binding proteins isolated from murine peritoneal macrophages are essentially identical to those homologous proteins previously isolated from P388D1 cells [20].
mesh_terms: D000727:Androgen-Binding Protein; D000818:Animals; D002352:Carrier Proteins / Q000302:isolation & purification; D007074:Immunoglobulin G / Q000378:metabolism*; D007700:Kinetics; D007941:Leukemia P388 / Q000276:immunology; D008264:Macrophages / Q000187:drug effects / Q000201:enzymology / Q000276:immunology*; D051379:Mice; D010713:Phosphatidylcholines / Q000378:metabolism; D049488:Phosphatidylethanolamine Binding Protein; D010740:Phospholipases / Q000378:metabolism*; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D049294:Phospholipid Transfer Proteins; D011961:Receptors, Fc / Q000302:isolation & purification / Q000378:metabolism*; D017452:Receptors, IgG; D011971:Receptors, Immunologic / Q000302:isolation & purification; D012397:Rosette Formation; D013864:Thioglycolates / Q000494:pharmacology*
chemical_list: D000727:Androgen-Binding Protein; D002352:Carrier Proteins; D007074:Immunoglobulin G; C511482:PEBP1 protein, human; D010713:Phosphatidylcholines; D049488:Phosphatidylethanolamine Binding Protein; D049294:Phospholipid Transfer Proteins; D011961:Receptors, Fc; D017452:Receptors, IgG; D011971:Receptors, Immunologic; D013864:Thioglycolates; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2