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Tn Antigen Is Expressed on Leukosialin from T-Lymphoid Cells¹

Department of Biotechnology, Faculty of Engineering, Kyoto Sangyo University, Kita-ku, Kyoto 603, Japan

² To whom requests for reprints should be addressed.

Various T-lymphoid cells were labeled with [³H] glucosamine and then cell lysates were prepared from them. The Tn antigen was immunoprecipitated and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by fluorography. The Tn antigen was found to be expressed on leukosialin, a major glycoprotein of T-lymphoid cells. The carbohydrate moieties of leukosialin were isolated from Jurkat and Molt 4 cells by alkaline borohydride treatment. The leukosialin in both cases predominantly contained single N-acetylgalactosamine residues, consistent with expression of the Tn antigen. Tryptic glycopeptides containing antigenic sites were isolated using an MLS 128 immunoaffinity column and purified by gel filtration and reverse phase column chromatographies. Sequence analyses revealed that all the glycopeptides obtained contained three consecutive residues of N-acetylgalactosamine-Ser/Thr, supporting the idea that the epitopic structure is a cluster of N-acetylgalactosamineSer/Thr.

¹ This work was supported in part by a Grant-in-Aid for Scientific Research and a Grant-in-Aid for Scientific Research on Priority Areas (05274107), from the Ministry of Education, Science and Culture of Japan; by the Fugaku Trust for Medicinal Research; and by a Grant-in-Aid from the Sankyo Foundation of Life Science.

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.

Received 8/ 6/93. Accepted 10/29/93.

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