Effects of enzymatic deglycosylation on the biological activities of human thrombin and antithrombin.
Sequential digestion of human thrombin and antithrombin with neuraminidase, beta-galactosidase, beta-N-acetylglucosaminidase, and endo-beta-N-acetylglucosaminidase D resulted in the successive removal of sialic acid, galactose, N-acetylglucosamine, and mannose and more N-acetylglucosamine residues. The products obtained after each stage of deglycosylation had electrophoretic mobilities that were consistent with the calculated change in mass expected from the cleavage of the sugar moieties. The modified thrombins did not lose fibrinogen-clotting activity, amidolytic activity, nor the ability to form complexes with antithrombin. In addition, asialothrombin and asialoagalactothrombin caused the same extent of platelet release as did control thrombin. The products obtained after removal of sugars from antithrombin retained thrombin-neutralizing activity. In the presence of heparin the inhibition of thrombin as well as factor Xa was enhanced. Thus, the sugar residues of thrombin and antithrombin are not required for the formation of enzyme-inhibitor complexes or for the other activities that were measured.
D000118:Acetylglucosaminidase / Q000378:metabolism; D000991:Antithrombins / Q000378:metabolism / Q000494:pharmacology*; D001792:Blood Platelets / Q000187:drug effects; D002241:Carbohydrates / Q000502:physiology*; D004151:Dipeptides / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006801:Humans; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D009439:Neuraminidase / Q000378:metabolism; D013329:Structure-Activity Relationship; D013917:Thrombin / Q000378:metabolism / Q000494:pharmacology*; D001616:beta-Galactosidase / Q000378:metabolism
D000991:Antithrombins; D002241:Carbohydrates; D004151:Dipeptides; C021129:S 2238; D005340:Fibrinogen; D006026:Glycoside Hydrolases; D009439:Neuraminidase; D001616:beta-Galactosidase; D000118:Acetylglucosaminidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D013917:Thrombin