Purification of cachectin, a lipoprotein lipase-suppressing hormone secreted by endotoxin-induced RAW 264.7 cells.
Previous studies have indicated that endotoxin and other bacterial and protozoal products can stimulate macrophages to produce a factor that can suppress the activity of the enzyme lipoprotein lipase (LPL), in vivo and in vitro. In the present report we describe the purification of this factor, cachectin, to apparent homogeneity from the conditioned medium of endotoxin-stimulated RAW 264.7 cells. The isolated protein has an isoelectric point of 4.7 and a subunit molecular weight of 17,000. Although cachectin's isoelectric point and molecular weight are similar to those described for interleukin 1, pure cachectin has no leukocyte-activating factor (LAF) activity. Cachectin at a concentration of 10(-11) M has the ability to suppress the LPL activity of the 3T3-L1 adipocyte cell line by 80%. Binding studies using radio-labeled cachectin and 3T3-L1 adipocytes and C2 myotubules revealed approximately 10(4) high-affinity receptors per cell on both cell types (Ka, 3 X 10(9]. Cachectin receptors were also present on liver membranes but were absent on erythrocytes and lymphocytes. The isolation of cachectin and characterization of its receptor should facilitate further investigations into the role of cachectin and other macrophage mediators in the metabolic derangements that occur during infection and cachexia.
D000273:Adipose Tissue / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004731:Endotoxins / Q000494:pharmacology; D007375:Interleukin-1 / Q000032:analysis; D007525:Isoelectric Focusing; D008071:Lipoprotein Lipase / Q000037:antagonists & inhibitors*; D008262:Macrophage Activation; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D008970:Molecular Weight; D009132:Muscles / Q000378:metabolism; D011506:Proteins / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha
D004731:Endotoxins; D007375:Interleukin-1; D011506:Proteins; D014409:Tumor Necrosis Factor-alpha; D008071:Lipoprotein Lipase