Abstract
Peptide-N4-(N-acetyl-β-glucosaminyl) asparagine amidase F (PNGase F) is an amidase/amidohydrolase that hydrolyzes the β-aspartylglycosylamine bond of asparagine-linked glycopeptides and glycoproteins. The enzyme recognizes mainly the polypeptide region around the glycosylation site and the inner core of the carbohydrate chain. All classes of oligosaccharides are released by PNGase F, making it the enzyme of choice for deglycosylation of most asparagine-linked glycans. Used in conjunction with current biotechnology, PNGase F is an important tool for structure/function analysis of glycoproteins, structural analysis of oligosaccharides, and biosynthetic studies of asparagine-linked oligosaccharides. PNGase F is available from several commercial sources, but is relatively expensive. Alternatively, the enzyme can be purified in large amounts by most laboratories with minimal work using the one-step protocol described herein.
