Glycan analysis of recombinant Aspergillus niger endo-polygalacturonase A.
The enzyme endo-polygalacturonase A, or PGA, is produced by the fungus, Aspergillus niger, and appears to play a critical role during invasion of plant cell walls. The enzyme has been homologously overexpressed in order to provide sufficient quantities of purified enzyme for structural and biological studies. We have characterized this enzyme in terms of its post-translational modifications (PTMs) and found it to be both N- and O-glycosylated. Additionally, we have characterized the glycosyl moieties using MALDI-TOF and LC-ESI mass spectrometry. The characterization of all PTMs on PGA, along with molecular modeling, allows us to reveal potential roles played by the glycans in modulating the interaction of the enzyme with other macromolecules.
Woosley
Bryan D
BD
Complex Carbohydrate Research Center, Department of Biochemistry and Molecular Biology, University of Georgia, 315 Riverbend Road, Athens, GA 30602-4712, USA.
Kim
Young Hwan
YH
Kumar Kolli
V S
VS
Wells
Lance
L
King
Dan
D
Poe
Ryan
R
Orlando
Ron
R
Bergmann
Carl
C
D000595:Amino Acid Sequence; D001234:Aspergillus niger / Q000737:chemistry*; D006020:Glycopeptides / Q000737:chemistry; D006031:Glycosylation; D008969:Molecular Sequence Data; D011096:Polygalacturonase / Q000737:chemistry* / Q000378:metabolism*; D011134:Polysaccharides / Q000737:chemistry*; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D006020:Glycopeptides; D011134:Polysaccharides; D011096:Polygalacturonase