Functional sialylated O-glycan to platelet aggregation on Aggrus (T1α/podoplanin) molecules expressed in Chinese hamster ovary cells

Mika Kaneko, Yukinari Kato, Akiko Kunita, Naoya Fujita, Takashi Tsuruo, Motoki Osawa

Research output: Contribution to journalArticlepeer-review

79 Citations (Scopus)

Abstract

Aggrus, also called T1α and podoplanin, is a novel platelet aggregation-inducing factor that is expressed in various carcinoma cells. Aggrus/T1α/podoplanin is known to be expressed in lung type I alveolar cells or lymphatic endothelial cells. However, its physiological role has not been clarified. To assess the attribution of glycosylation to Aggrus platelet aggregation activity, recombinant molecules were stably expressed in a series of Chinese hamster ovary (CHO) cell mutants, N-glycan-deficient Lec1, CMP-sialic acid transporter-deficient Lec2, and UDP-galactose transporter-deficient Lec8. A new anti-human Aggrus monoclonal antibody, YM-1, was established to detect the expression of human Aggrus on these CHO cell mutants. Aggrus on Lec1 cells induced platelet aggregation, but those on Lec2 and Lec8 cells did not. Further, the glycans on Aggrus were analyzed by lectin blotting. Aggrus expressed in CHO and Lec1 cells showed Wheat-germ agglutinin, Jacalin, and Vicia villosa lectin bindings. Lectin blotting results indicated that sialylated core 1 structures, sialic acid plus Galβ1,3GalNAc-Ser/Thr, were critical for the platelet aggregation activity. This oligosaccharide structure is known as tumor-associated antigen, which is potentially related to the metastasis process of cancer cells.

Original languageEnglish
Pages (from-to)38838-38843
Number of pages6
JournalJournal of Biological Chemistry
Volume279
Issue number37
DOIs
Publication statusPublished - 2004 Sep 10
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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