Elucidation of N-glycosylation sites on human platelet proteins: a glycoproteomic approach.

Article Details

Citation

Lewandrowski U, Moebius J, Walter U, Sickmann A

Elucidation of N-glycosylation sites on human platelet proteins: a glycoproteomic approach.

Mol Cell Proteomics. 2006 Feb;5(2):226-33. Epub 2005 Oct 31.

PubMed ID
16263699 [ View in PubMed
]
Abstract

Among known platelet proteins, a prominent and functionally important group is represented by glycoprotein isoforms. They account e.g. for secretory proteins and plasma membrane receptors including integrins and glycoprotein VI as well as intracellular components of cytosol and organelles including storage proteins (multimerin 1 etc.). Although many of those proteins have been studied for some time with regard to their function, little attention has been paid with respect to their glycosylation sites. Here we report the analysis of N-glycosylation sites of human platelet proteins. For the enrichment of glycopeptides, lectin affinity chromatography as well as chemical trapping of protein bound oligosaccharides was used. Therefore, concanavalin A was used for specific interaction with carbohydrate species along with periodic acid oxidation and hydrazide bead trapping of glycosylated proteins. Derivatization by peptide:N-glycosidase F yielded deglycosylated peptides, which provided the basis for the elucidation of proteins and their sites of modification. Using both methods in combination with nano-LC-ESI-MS/MS analysis 70 different glycosylation sites within 41 different proteins were identified. Comparison with the Swiss-Prot database established that the majority of these 70 sites have not been specifically determined by previous research projects. With this approach including hydrazide bead affinity trapping, the immunoglobulin receptor G6f, which is known to couple to the Ras-mitogen-activated protein kinase pathway in the immune system, was shown here for the first time to be present in human platelets.

DrugBank Data that Cites this Article

Polypeptides
NameUniProt ID
Coagulation factor VP12259Details
Antithrombin-IIIP01008Details
Cathepsin DP07339Details
Alpha-1-antitrypsinP01009Details
Transforming growth factor beta-1P01137Details
Platelet glycoprotein Ib alpha chainP07359Details
HeparanaseQ9Y251Details
Integrin beta-3P05106Details
P-selectinP16109Details
EndoplasminP14625Details
Fibrinogen beta chainP02675Details
Fibrinogen gamma chainP02679Details
Disintegrin and metalloproteinase domain-containing protein 10O14672Details
Complement C3P01024Details
Endothelin-converting enzyme 1P42892Details
ERO1-like protein betaQ86YB8Details
Lysosomal protective proteinP10619Details
Complement component C9P02748Details
ClusterinP10909Details
Beta-galactosidaseP16278Details