Elucidation of N-glycosylation sites on human platelet proteins: a glycoproteomic approach.
Article Details
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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
Name UniProt ID Coagulation factor V P12259 Details Antithrombin-III P01008 Details Cathepsin D P07339 Details Alpha-1-antitrypsin P01009 Details Transforming growth factor beta-1 proprotein P01137 Details Platelet glycoprotein Ib alpha chain P07359 Details Heparanase Q9Y251 Details Integrin beta-3 P05106 Details P-selectin P16109 Details Endoplasmin P14625 Details Fibrinogen beta chain P02675 Details Fibrinogen gamma chain P02679 Details Disintegrin and metalloproteinase domain-containing protein 10 O14672 Details Complement C3 P01024 Details Endothelin-converting enzyme 1 P42892 Details ERO1-like protein beta Q86YB8 Details Lysosomal protective protein P10619 Details Complement component C9 P02748 Details Clusterin P10909 Details Beta-galactosidase P16278 Details