Enzymatic deglycosylation of glycoproteins

Min Sung Kim, Dan Leahy

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Recombinant protein expression using eukaryotic expression systems has certain advantages, such as addition of posttranslational modifications that help protein stability and activity. Asparagine-linked sugar attachment is one of the most common posttranslation modifications. However, sugar modification can impede the growth of high-quality protein crystals for structural studies using X-ray crystallography. To overcome this problem, consensus sites of N-linked attachments can be mutated into other similar residues, such as aspartic acid. Alternatively, enzymatic deglycosylation can be used to remove sugars. Peptide-N-Glycosidase F (PNGase F; EC 3.5.1.52) and Endoglycosidase H (Endo H; EC 3.2.1.96) are the most popular enzymes for this purpose.

Original languageEnglish (US)
Title of host publicationLaboratory Methods in Enzymology
Subtitle of host publicationCell, Lipid and Carbohydrate
PublisherAcademic Press Inc.
Pages259-263
Number of pages5
ISBN (Print)9780124200678
DOIs
StatePublished - Jan 1 2013

Publication series

NameMethods in Enzymology
Volume533
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • Affinity purification methods
  • Enzymatic deglycosylation
  • Glycoproteins
  • Maltose-binding protein (MBP)
  • Oligosaccharides
  • Recombinant proteins

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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