Identification and quantification of N-linked glycoproteins using hydrazide chemistry, stable isotope labeling and mass spectrometry.
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Zhang H, Li XJ, Martin DB, Aebersold R
Identification and quantification of N-linked glycoproteins using hydrazide chemistry, stable isotope labeling and mass spectrometry.
Nat Biotechnol. 2003 Jun;21(6):660-6. Epub 2003 May 18.
- PubMed ID
- 12754519 [ View in PubMed]
- Abstract
Quantitative proteome profiling using stable isotope protein tagging and automated tandem mass spectrometry (MS/MS) is an emerging technology with great potential for the functional analysis of biological systems and for the detection of clinical diagnostic or prognostic marker proteins. Owing to the enormous complexity of proteomes, their comprehensive analysis is an as-yet-unresolved technical challenge. However, biologically or clinically important information can be obtained if specific, information-rich protein classes, or sub-proteomes, are isolated and analyzed. Glycosylation is the most common post-translational modification. Here we describe a method for the selective isolation, identification and quantification of peptides that contain N-linked carbohydrates. It is based on the conjugation of glycoproteins to a solid support using hydrazide chemistry, stable isotope labeling of glycopeptides and the specific release of formerly N-linked glycosylated peptides via peptide- N-glycosidase F (PNGase F). The recovered peptides are then identified and quantified by MS/MS. We applied the approach to the analysis of plasma membrane proteins and proteins contained in human blood serum.
DrugBank Data that Cites this Article
- Polypeptides
Name UniProt ID Neprilysin P08473 Details Low-density lipoprotein receptor P01130 Details Glutamate carboxypeptidase 2 Q04609 Details Lysosomal alpha-glucosidase P10253 Details Alpha-2-macroglobulin P01023 Details Alpha-1-acid glycoprotein 1 P02763 Details Cathepsin D P07339 Details Alpha-1-antitrypsin P01009 Details Beta-glucuronidase P08236 Details Lysosomal acid glucosylceramidase P04062 Details Kininogen-1 P01042 Details Basement membrane-specific heparan sulfate proteoglycan core protein P98160 Details Beta-hexosaminidase subunit beta P07686 Details Endoplasmin P14625 Details Laminin subunit gamma-1 P11047 Details Procathepsin L P07711 Details Plasma kallikrein P03952 Details Plasma protease C1 inhibitor P05155 Details Complement C3 P01024 Details Integrin alpha-V P06756 Details Cation-independent mannose-6-phosphate receptor P11717 Details Retinoid-inducible serine carboxypeptidase Q9HB40 Details Palmitoyl-protein thioesterase 1 P50897 Details Carboxypeptidase D O75976 Details N-acetylmuramoyl-L-alanine amidase Q96PD5 Details Complement C4-A P0C0L4 Details Complement C4-B P0C0L5 Details Sodium/potassium-transporting ATPase subunit beta-1 P05026 Details Sodium/potassium-transporting ATPase subunit beta-3 P54709 Details Alpha-2-HS-glycoprotein P02765 Details Clusterin P10909 Details Inter-alpha-trypsin inhibitor heavy chain H2 P19823 Details Kallistatin P29622 Details Inter-alpha-trypsin inhibitor heavy chain H1 P19827 Details Inter-alpha-trypsin inhibitor heavy chain H4 Q14624 Details Alpha-1-antichymotrypsin P01011 Details N-sulphoglucosamine sulphohydrolase P51688 Details Haptoglobin P00738 Details