Enzymatic modifications of human fibroblast and leukocyte interferons.
The sensitivity of highly purified human fibroblast interferon and partially purified human leukocyte interferon to several proteolytic and glycolytic enzymes was determined with respect to antiviral activity, isoelectric point, molecular weight, and thermal stability. Leucine aminopeptidase altered the distribution of isoelectric points for both interferons but produced little change in molecular weights; this enzyme somewhat reduced the activity of only leukocyte interferon. Treatment of fibroblast interferon with carboxypeptidases A and B did not greatly decrease antiviral activity, but it did slightly reduce the molecular weight of the interferon and substantially altered the distribution of isoelectric point values; similar treatment of leukocyte interferon caused some loss in activity, especially of the 17,000-molecular-weight species. Both interferons were inactivated rapidly by treatment with the endoproteases trypsin, pepsin, bromelain, and subtilisin. Chymotrypsin shifted the isoelectric points of both interferons, but only leukocyte interferon was significantly inactivated. Treatment with neuraminidase and beta-galactosidase changed the isoelectric point distribution but did not affect the activity or thermal stability of either interferon; such a treatment reduced the molecular weight of fibroblast interferon and the size heterogeneity of leukocyte interferon. Treatment with neuraminidase and then leucine aminopeptidase greatly reduced the activity of both interferons, especially leukocyte interferon. The data indicate that biologically active forms of fibroblast and leukocyte interferons can be distinguished by their relative sensitivity to certain proteases.
D002268:Carboxypeptidases / Q000378:metabolism; D002918:Chymotrypsin / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D006358:Hot Temperature; D006801:Humans; D007372:Interferons / Q000378:metabolism*; D007526:Isoelectric Point; D007931:Leucyl Aminopeptidase / Q000378:metabolism; D007962:Leukocytes / Q000378:metabolism*; D008970:Molecular Weight; D009439:Neuraminidase / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
D007372:Interferons; D009439:Neuraminidase; D001616:beta-Galactosidase; D002268:Carboxypeptidases; D007931:Leucyl Aminopeptidase; D002918:Chymotrypsin