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Characterization of endopolygalacturonase (EC 3.2.1.15) from Aspergillus niger as glycoprotein by electrophoretic methods and lectin affino‐blotting
Author(s) -
Raab Barbara
Publication year - 1992
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.11501301177
Subject(s) - isoelectric focusing , concanavalin a , lectin , glycoprotein , mannose , biochemistry , chemistry , aspergillus niger , gel electrophoresis , glycosylation , polyacrylamide gel electrophoresis , electrophoresis , chromatography , enzyme , in vitro
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.