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J. Biol. Chem., Vol. 279, Issue 37, 38838-38843, September 10, 2004

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Functional Sialylated O-Glycan to Platelet Aggregation on Aggrus (T1/Podoplanin) Molecules Expressed in Chinese Hamster Ovary Cells^*

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

From the Department of Experimental and Forensic Pathology, Yamagata University School of Medicine, 2-2-2, Iida-nishi, Yamagata 990-9585, Japan, the Institute of Molecular and Cellular Biosciences, The University of Tokyo, 1-1-1, Yayoi, Bunkyo-ku, Tokyo 113-0032 Japan, and the ^¶Cancer Chemotherapy Center, Japanese Foundation for Cancer Research, 1-37-1, Kami-Ikebukuro, Toshima-ku, Tokyo 170-8455, Japan

Aggrus, also called T1 and podoplanin, is a novel platelet aggregation-inducing factor that is expressed in various carcinoma cells. Aggrus/T1/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 Gal1,3GalNAc-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.

Received for publication, June 28, 2004

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^|| To whom correspondence should be addressed. Tel.: 81-23-628-5271; Fax: 81-23-628-5273; E-mail: mosawa{at}med.id.yamagata-u.ac.jp.

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