Some physicochemical properties of human leucocyte migration inhibitory factor (LIF).
Leucocyte migration inhibitory factor (LIF) obtained from human lymphocytes stimulated with concanavalin A was consistently and irreversibly blocked by the serine-esterase inhibitor phenyl-methyl sulphonylfuoride (PMSF). This effect was not due to fluoride ions, hydrolysis products of PMSF or to impurities. PMSF pulse treatment of human buffy coat cells did not affect cell migration under agarose. LIF was also irreversibly destroyed by treatment with L-cysteine and 2-mercapto-ethanol, suggesting that the molecule contains disulphide linkage groups decisive for its configuration and biological activity. Di-sodium EDTA completely inhibited LIF activity but only if present during the entire migration period. Removal of EDTA before LIF assay restored LIF activity. Leucotye migration was neighter influenced by L-cysteine nor by EDTA. LIF activity was slightly diminished after treatment at 56 degrees C for 1 h and completely lost at 80 degrees C for 1/2 h. Furthermore, LIF appeared rather stable when treated at pH values between 4 and 11. These findings suggest, but do not prove, an esterase or a protease nature of human LIF.
D003208:Concanavalin A / Q000494:pharmacology; D003545:Cysteine / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007962:Leukocytes / Q000276:immunology*; D008213:Lymphocyte Activation / Q000187:drug effects; D008263:Macrophage Migration-Inhibitory Factors / Q000032:analysis / Q000378:metabolism; D008623:Mercaptoethanol / Q000494:pharmacology; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology*; D013450:Sulfones / Q000494:pharmacology*; D013696:Temperature
D008263:Macrophage Migration-Inhibitory Factors; D013450:Sulfones; D003208:Concanavalin A; D010664:Phenylmethylsulfonyl Fluoride; D008623:Mercaptoethanol; D004492:Edetic Acid; D003545:Cysteine