Lysosomal beta-hexosaminidase is highly resistant towards proteolytic degradation in vitro.
1. A partially purified enzyme preparation of beta-hexosaminidase from human fibroblasts was treated with proteases and the effect on its molecular weight and enzymatic activity was studied. 2. Both the forms A and B of the enzyme appeared to be resistant to a protease treatment that degraded the majority of the contaminating proteins to a large extent. 3. The same result was obtained with enzyme preparations from cells treated with tunicamycin. 4. Also the molecular weights of the individual polypeptide chains of the enzyme were not decreased, as was shown by SDS-PAGE, followed by immuno-blotting.
Overdijk
B
B
Department of Medical Chemistry, Faculty of Medicine, Vrije Universiteit, Amsterdam, The Netherlands.
Beem
E P
EP
Van der Wal
C J
CJ
Jongenelen
C A
CA
D002478:Cells, Cultured; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D008247:Lysosomes / Q000201:enzymology*; D008970:Molecular Weight; D011402:Pronase / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism; D014415:Tunicamycin / Q000494:pharmacology; D001619:beta-N-Acetylhexosaminidases / Q000096:biosynthesis / Q000187:drug effects / Q000378:metabolism*
D014415:Tunicamycin; D001619:beta-N-Acetylhexosaminidases; D014357:Trypsin; D011402:Pronase