Human leukocyte migration inhibitory factor (LIF) III. Further investigations on the serine protease nature of this lymphokine and its preference for arginine amides.
Previous findings suggesting an esterase and protease nature of human leukocyte migration inhibitory factor (LIF) were extended by testing the ability of different protease and esterase inhibitors to reduce LIF activity. The serine-specific inhibitors phenylmethylsulfonyl fluoride (PMSF) and physostigmine (eserine) markedly reduced LIF activity, whereas the histidine-specific inhibitors N-tosyl-L-lysine chloromethyl ketone (TLCK) and L-tosylamide-2-phenylethyl chloromethyl ketone (TPCK) were inactive. That LIF might act as an esterase and a protease was further strengthened by the ability of pralidoxime methansulfonate (2-PAM) to reestablish LIF activity of supernatants previously inactivated by PMSF. Furthermore, the arginine amides N-benzoyl-L-arginine-p-nitroanilide (BApNA) and N-benzoyl-L-phenylalanyl-L-valyl-L-arginine-p-nitroanilide (BPVApNA) were shown to satisfy the substrate specificities of the putative LIF enzyme. Indeed, BPVApNA seemed to possess a particularly strong affinity for LIF, indicating its potential role in an enzymatic LIF assay.
D000577:Amides / Q000494:pharmacology*; D001120:Arginine / Q000494:pharmacology*; D006801:Humans; D007962:Leukocytes / Q000276:immunology*; D008263:Macrophage Migration-Inhibitory Factors; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D010830:Physostigmine / Q000494:pharmacology; D011220:Pralidoxime Compounds / Q000494:pharmacology; D014107:Tosyllysine Chloromethyl Ketone / Q000494:pharmacology; D014108:Tosylphenylalanyl Chloromethyl Ketone / Q000494:pharmacology
D000577:Amides; D008263:Macrophage Migration-Inhibitory Factors; D011220:Pralidoxime Compounds; D014107:Tosyllysine Chloromethyl Ketone; D014108:Tosylphenylalanyl Chloromethyl Ketone; D010664:Phenylmethylsulfonyl Fluoride; D001120:Arginine; D010830:Physostigmine