Characterization of endopolygalacturonase (EC 3.2.1.15) from Aspergillus niger as glycoprotein by electrophoretic methods and lectin affino-blotting.
Chromatographically purified endopolygalacturonase (PG) from Aspergillus niger was deglycosylated with N-glycosidase F (PNGase F) and characterized by means of sodium dodecyl sulfate (SDS)-electrophoresis, polyacrylamide gel electrophoresis (PAGE) without denaturing agents, isoelectric focusing (IEF) and lectin affino-blotting. The results show that PG, which is apparently homogeneous in SDS-PAGE but heterogeneous in IEF and PAGE, consists of at least two polypeptide chains with different glycosylation patterns. The component with the higher electrophoretic mobility is deglycosylated with PNGase F and reacts with concanavalin A (Con A) and Galanthus nivalis agglutinin (GNA), indicating a "high mannose" or "hybrid"-type of glycoprotein (GP). The other component may contain O-glycosidically linked mannose, N-acetylglucosamine or glucose.
Raab
B
B
Deutsches Institut für Ernhrungsforschung Potsdam-Rehbrücke, Germany.
D001234:Aspergillus niger / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D027801:Galanthus; D006023:Glycoproteins / Q000737:chemistry; D006031:Glycosylation; D015151:Immunoblotting; D007525:Isoelectric Focusing; D037102:Lectins; D037121:Plant Lectins; D011096:Polygalacturonase / Q000737:chemistry*
D006023:Glycoproteins; D037102:Lectins; D037121:Plant Lectins; D011096:Polygalacturonase