PNGase F catalyzes de-N-glycosylation in a domestic microwave

Anal Biochem. 2012 Aug 1;427(1):33-5. doi: 10.1016/j.ab.2012.04.011. Epub 2012 Apr 16.

Abstract

Common de-N-glycosylation protocols usually require a lengthy incubation time. Although pressure cycling technology or scientific microwave reactors can accelerate this enzyme reaction, they may not be easily accessible. In this brief report, we employed an alternative strategy using a standard domestic microwave oven to perform the de-N-glycosylation. Model glycoproteins (bovine RNase B, bovine fetuin, and human IgG) and a complex mixture from human plasma were fully deglycosylated in 20 min, without any apparent adverse affects on the glycans or protein backbones. This new method provides a simple and inexpensive solution to achieve rapid de-N-glycosylation.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Catalysis
  • Cattle
  • Fetuins / analysis
  • Glycoproteins / chemistry
  • Glycosylation / radiation effects*
  • Humans
  • Immunoglobulin G / analysis
  • Microwaves*
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / chemistry*
  • Polysaccharides / chemistry
  • Ribonuclease, Pancreatic / analysis

Substances

  • Fetuins
  • Glycoproteins
  • Immunoglobulin G
  • Polysaccharides
  • Ribonuclease, Pancreatic
  • pancreatic ribonuclease B, bovine
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase