PNGase F (Glycopeptidase F)
Glycopeptidase F, also known as PNGase F or Peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine amidase, removes N-glycan groups from glycoproteins and glycopeptides. Deglycosylation enables proteomic analysis, including MALDI-TOF mass spectrometry. PNGase F is often used to determine if proteins contain glycosylated peptides and if deglycosylation affects a protein's enzymatic activity. Removal of N-linked glycans prior to MALDI-TOF and other mass spectrometry procedures may also be performed to reduce protein heterogeneity and thereby facilitate proteomic analysis. PNGase F is one of the most widely used enzymes for deglycosylation of glycoproteins and glycopeptides.
More Information
Applications
- Determination of protein and/or peptide glycosylation level
- Analysis of N-glycan functional roles
- Reduction of protein heterogeneity
- MALDI-TOF or other mass spectrometry
Components
- Denaturing buffer: 1% SDS / 1M Tris-HCl (pH 8.6; 500 μl)
- Native buffer: 1M Tris-HCl (pH 8.6; 500 μl)
- Stabilized solution: 5% NP-40 (500 μl)
- Control glycoprotein: 10 mg/ml Bovine Fetuin (10 μl)
Source
Recombinant Escherichia coli encoding PNGase F.
Storage
−20°C
Substrate specificity
Specific cleavage of N-glycan (GluNAc-Asn) bonds in glycoproteins and glycopeptides.
PNGase F (Glycopeptidase F) will not cleave N-glycan groups when the innermost GlcNAc residue is linked to an alpha1-3 Fucose residue. This modification is found in plant glycoproteins as well as some insect glycoproteins.
Definition of activity
One unit is the amount of enzyme required to hydrolyze 1 μmol of dansyl fetuin glycopeptide within 1 minute at 37°C at pH 8.6.
Properties
- Molecular weight: approx. 36.5 kDa (SDS-PAGE)
Note: Depending on proteolytic activity of host bacteria, recombinant PNGase F may have 15 or 18 additional amino acid residues attached to its N-terminus. However, PNGase F glycosylase activity has been confirmed to be identical to that of native PNGase F via experiments using RNase B and Fetuin as substrates.
- Optimum pH: 8.6
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
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