Influence of carbohydrate side chains on activity of tissue-type plasminogen activator.
When messenger RNA (mRNA) from both untreated and phorbol ester-treated melanoma cells is translated in simple reticulocyte lysates, tissue-type plasminogen activator can be immunoprecipitated by an affinity-purified antibody as a approximately 52,000 mol wt protein, with no detectable biological (plasminogen activating) activity. When the reticulocyte lysate system is supplemented with a preparation of microsomal membranes, biological activity becomes detectable and a 63,000 mol wt protein can be immunoprecipitated with the same antibody. Furthermore, when natural tissue-type plasminogen activator (mol wt approximately equal to 70,000) is incubated with different glycosidases, distinct alterations in the electrophoretic mobility of the molecules are observed, together with alterations in the level of biological activity. While treatment with neuraminidase and beta-galactosidase caused decreases in activity, alpha-mannosidase caused an increase. These results suggest that the carbohydrate part of the molecule can influence its biological behavior.
D000577:Amides / Q000378:metabolism; D002478:Cells, Cultured; D005337:Fibrin / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008361:Mannosidases / Q000378:metabolism; D008861:Microsomes / Q000378:metabolism; D008970:Molecular Weight; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D012156:Reticulocytes / Q000378:metabolism; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D010959:Tissue Plasminogen Activator / Q000378:metabolism; D014415:Tunicamycin / Q000494:pharmacology; D043323:alpha-Mannosidase
D000577:Amides; D006023:Glycoproteins; D014415:Tunicamycin; D005337:Fibrin; D005340:Fibrinogen; D008361:Mannosidases; D043323:alpha-Mannosidase; D010959:Tissue Plasminogen Activator