title: Enzymatic deglycosylation of glycoproteins.
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.
authors.affiliation: Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Min-Sung
authors.initials: MS
authors.lastname: Kim
authors.forename: Dan
authors.initials: D
authors.lastname: Leahy
mesh_terms: D000818:Animals; D016466:CHO Cells; D003412:Cricetulus; D018360:Crystallography, X-Ray; D006023:Glycoproteins / Q000737:chemistry* / Q000378:metabolism*; D006031:Glycosylation; D057809:HEK293 Cells; D006801:Humans; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000378:metabolism; D015202:Protein Engineering / Q000295:instrumentation / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D006023:Glycoproteins; D011994:Recombinant Proteins; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase
keywords: Affinity purification methods; Enzymatic deglycosylation; Glycoproteins; Maltose-binding protein (MBP); Oligosaccharides; Recombinant proteins