Publications

• N-glycoproteomics in plants: perspectives and challenges.

  Song W., Henquet M.G., Mentink R.A., van Dijk A.J., Cordewener J.H., Bosch D., America A.H., van der Krol A.R.

  In eukaryotes, proteins that are secreted into the ER are mostly modified by N-glycans on consensus NxS/T sites. The N-linked glycan subsequently undergoes varying degrees of processing by enzymes which are spatially distributed over the ER and the Golgi apparatus. The post-ER N-glycan processing ... >> More

  In eukaryotes, proteins that are secreted into the ER are mostly modified by N-glycans on consensus NxS/T sites. The N-linked glycan subsequently undergoes varying degrees of processing by enzymes which are spatially distributed over the ER and the Golgi apparatus. The post-ER N-glycan processing to complex glycans differs between animals and plants, with consequences for N-glycan and glycopeptide isolation and characterization of plant glycoproteins. Here we describe some recent developments in plant glycoproteomics and illustrate how general and plant specific technologies may be used to address different important biological questions. << Less

  J Proteomics 74:1463-1474(2011) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Dolichyldiphosphoryloligosaccharide--protein oligosaccharyltransferase; solubilization, purification, and properties.

  Das R.C., Heath E.C.

  Dolichyldiphosphoryloligosaccharide-protein oligosaccharyltransferase was solubilized from hen oviduct rough endoplasmic reticulum by extraction with 0.2% Nonidet P40. Oligosaccharyltransferase activity was assayed in an incubation mixture containing Glc(n)-Man(x)-GlcNAc(2)-diphosphoryldolichol as ... >> More

  Dolichyldiphosphoryloligosaccharide-protein oligosaccharyltransferase was solubilized from hen oviduct rough endoplasmic reticulum by extraction with 0.2% Nonidet P40. Oligosaccharyltransferase activity was assayed in an incubation mixture containing Glc(n)-Man(x)-GlcNAc(2)-diphosphoryldolichol as an oligosaccharyl donor and the (125)I-labeled tryptic peptide consisting of residues 29-58 from bovine alpha-lactalbumin as acceptor. The transferase was purified approximately 2000-fold by fractionation on a bovine alpha-lactalbumin-Sepharose column; the active material bound quantitatively to the gel and was eluted by removal of divalent cation from the wash buffer. The product of the transferase activity, (125)I-glycopeptide, was determined as concanavalin A-agarose-adsorbed radioactivity by a filter disc assay method. (125)I-Labeled concanavalin A-agarose-bound product was characterized as a glycopeptide as follows: (i) gel filtration behavior on Sephadex G-50; (ii) elution from concanavalin A-agarose with 1% alpha-methyl mannoside; (iii) absence of affinity for ricin-Sepharose and loss of affinity for concanavalin A-agarose after treatment with endo-beta-N-acetylglucosaminidase H; (iv) enzymatic synthesis of identical product upon using [(3)H]oligosaccharyldiphosphoryldolichol and unlabeled peptide acceptor; and (v) digestion of (3)H-labeled peptide with Pronase, resulting in the formation of lower molecular weight glycopeptide. Oligosaccharyltransferase activity exhibited an absolute requirement for divalent cations (3 mM Mn(2+); Mg(2+) was 30% as effective), complete dependence on exogenously supplied peptide acceptor (1.33 mug/ml) and oligosaccharyldiphosphoryldolichol (approximately 10 nmol/ml), and an optimum pH between 7 and 7.5. << Less

  Proc Natl Acad Sci U S A 77:3811-3815(1980) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.