Functional sialylated O-glycan to platelet aggregation on Aggrus (T1alpha/Podoplanin) molecules expressed in Chinese hamster ovary cells.
Aggrus, also called T1alpha and podoplanin, is a novel platelet aggregation-inducing factor that is expressed in various carcinoma cells. Aggrus/T1alpha/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 Galbeta1,3GalNAc-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.
Kaneko
Mika
M
Department of Experimental and Forensic Pathology, Yamagata University School of Medicine, 2-2-2, Iida-nishi, Yamagata 990-9585, Japan.
Kato
Yukinari
Y
Kunita
Akiko
A
Fujita
Naoya
N
Tsuruo
Takashi
T
Osawa
Motoki
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D015153:Blotting, Western; D016466:CHO Cells; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D007150:Immunohistochemistry; D037102:Lectins / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000502:physiology*; D008565:Membrane Proteins / Q000502:physiology*; D008954:Models, Biological; D009154:Mutation; D009844:Oligosaccharides / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D010974:Platelet Aggregation; D011134:Polysaccharides / Q000737:chemistry*; D011233:Precipitin Tests; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000378:metabolism; D012794:Sialic Acids / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D000098853:Podoplanin
D000911:Antibodies, Monoclonal; D037102:Lectins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D009844:Oligosaccharides; D010455:Peptides; D011134:Polysaccharides; D011994:Recombinant Proteins; D012794:Sialic Acids; C117468:PDPN protein, human; D000098853:Podoplanin; C092025:Pdpn protein, rat