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
NameUniProt ID
NeprilysinP08473Details
Low-density lipoprotein receptorP01130Details
Glutamate carboxypeptidase 2Q04609Details
Lysosomal alpha-glucosidaseP10253Details
Alpha-2-macroglobulinP01023Details
Alpha-1-acid glycoprotein 1P02763Details
Cathepsin DP07339Details
Alpha-1-antitrypsinP01009Details
Beta-glucuronidaseP08236Details
GlucosylceramidaseP04062Details
Kininogen-1P01042Details
Basement membrane-specific heparan sulfate proteoglycan core proteinP98160Details
Beta-hexosaminidase subunit betaP07686Details
EndoplasminP14625Details
Laminin subunit gamma-1P11047Details
Cathepsin L1P07711Details
Plasma kallikreinP03952Details
Plasma protease C1 inhibitorP05155Details
Complement C3P01024Details
Integrin alpha-VP06756Details
Cation-independent mannose-6-phosphate receptorP11717Details
Retinoid-inducible serine carboxypeptidaseQ9HB40Details
Palmitoyl-protein thioesterase 1P50897Details
Carboxypeptidase DO75976Details
N-acetylmuramoyl-L-alanine amidaseQ96PD5Details
Complement C4-AP0C0L4Details
Complement C4-BP0C0L5Details
Sodium/potassium-transporting ATPase subunit beta-1P05026Details
Sodium/potassium-transporting ATPase subunit beta-3P54709Details
Alpha-2-HS-glycoproteinP02765Details
ClusterinP10909Details
Inter-alpha-trypsin inhibitor heavy chain H2P19823Details
KallistatinP29622Details
Inter-alpha-trypsin inhibitor heavy chain H1P19827Details
Inter-alpha-trypsin inhibitor heavy chain H4Q14624Details
Alpha-1-antichymotrypsinP01011Details
N-sulphoglucosamine sulphohydrolaseP51688Details
HaptoglobinP00738Details