Abstract
Podoplanin (Aggrus) is a mucin-type sialoglycoprotein that plays a key role in tumor cell-induced platelet aggregation. Podoplanin possesses a platelet aggregation-stimulating (PLAG) domain, and Thr52 in the PLAG domain of human podoplanin is important for its activity. Endogenous or recombinant human podoplanin were purified, and total glycosylation profiles were surveyed by lectin microarray. Analyses of glycopeptides produced by Edman degradation and mass spectrometry revealed that the disialyl-corel (NeuAc alpha2-3Gal beta l-3(NeuAc alpha2-6)GalNAc alpha l-O-Thr) structure was primarily attached to a glycosylation site at residue Thr52. Sialic acid-deficient podoplanin recovered its activity after additional sialylation. These results indicated that the sialylated Corel at Thr52 is critical for podoplanin-induced platelet aggregation.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Blood Platelets / drug effects
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CHO Cells
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Carbohydrate Sequence
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Cricetinae
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Cricetulus
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Glycosylation
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Humans
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Lectins / chemistry
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Mass Spectrometry
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Membrane Glycoproteins / chemistry*
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Membrane Glycoproteins / isolation & purification
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Membrane Glycoproteins / pharmacology
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Molecular Sequence Data
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N-Acetylneuraminic Acid / analysis
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Organophosphorus Compounds / chemistry
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Peptide Fragments / chemistry
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Platelet Aggregation
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Polysaccharides / analysis
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Polysaccharides / chemistry*
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Protein Array Analysis
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Recombinant Proteins / chemistry
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / pharmacology
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Threonine / chemistry
Substances
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2-(4-isothiocyanatophenoxy)-1,3,2-dioxaphosphinene 2-oxide
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Lectins
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Membrane Glycoproteins
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Organophosphorus Compounds
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PDPN protein, human
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Peptide Fragments
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Polysaccharides
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Recombinant Proteins
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Threonine
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N-Acetylneuraminic Acid