title: PNGase F catalyzes de-N-glycosylation in a domestic microwave.
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.
authors.affiliation: Department of Urology and The Proteomics Center, Children's Hospital Boston and Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhou
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Briscoe
authors.forename: John W
authors.initials: JW
authors.lastname: Froehlich
authors.forename: Richard S
authors.initials: RS
authors.lastname: Lee
mesh_terms: D000818:Animals; D002384:Catalysis; D002417:Cattle; D060748:Fetuins / Q000032:analysis; D006023:Glycoproteins / Q000737:chemistry; D006031:Glycosylation / Q000528:radiation effects*; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D008872:Microwaves; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000737:chemistry*; D011134:Polysaccharides / Q000737:chemistry; D012259:Ribonuclease, Pancreatic / Q000032:analysis
chemical_list: D060748:Fetuins; D006023:Glycoproteins; D007074:Immunoglobulin G; D011134:Polysaccharides; D012259:Ribonuclease, Pancreatic; C504335:pancreatic ribonuclease B, bovine; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase