Physicochemical characterization of human fibroblast migration inhibitory factor.
We recently reported a new lymphokine activity that affects fibroblasts by inhibiting their spontaneous migration. Human fibroblast migration inhibitory factor (FIF) obtained from concanavalin A (Con A)-stimulated human lymphocytes was characterized by Sephadex gel filtration and by enzyme treatment. FIF was found to be stable at 56 degrees C for 15 min but destroyed at 80 degrees C or at pH lower than 5. Gel filtration revealed two peaks of FIF activity 15,000 and at 34,000 Da. FIF activity was lost following treatment with trypsin, chymotrypsin, and neuraminidase and FIF could not be generated in the presence of inhibitors of glycosylation, suggesting that the molecule was a glycoprotein. FIF could be removed by adsorption to human fibroblasts but not to PMN, monocytes, or red blood cells. Further studies were carried out to investigate the role of sugars in the interaction of FIF with the target cells. Human FIF activity was significantly reduced in the presence of several sugars including alpha-methyl-D-mannoside, L-xylose, N-acetyl-D-glucosamine, D-mannose, L-rhamnose but not L-fucose. Preincubation of human fibroblasts with alpha-methyl-D-mannoside prevented their response to FIF. In contrast, pretreatment of fibroblasts with mannosidase had no effect, suggesting that alpha-methyl-D-mannoside was an essential component of the FIF molecule recognized by the FIF receptor on fibroblasts.
D002464:Cell Migration Inhibition; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002918:Chymotrypsin / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008222:Lymphokines / Q000032:analysis*; D008361:Mannosidases / Q000378:metabolism; D008362:Mannosides / Q000494:pharmacology; D008970:Molecular Weight; D009005:Monosaccharides / Q000494:pharmacology; D009439:Neuraminidase / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D013696:Temperature; D014357:Trypsin / Q000378:metabolism; D014415:Tunicamycin / Q000494:pharmacology
D008222:Lymphokines; D008362:Mannosides; D009005:Monosaccharides; C037155:fibroblast migration inhibitory factor; D014415:Tunicamycin; D008361:Mannosidases; D009439:Neuraminidase; D002918:Chymotrypsin; D014357:Trypsin