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J. Biochem, 1995, Vol. 118, No. 3 658-664
© 1995 Japanese Biochemical Society

other

Molecular Cloning and Characterization of a Third Type of N-Glycan {alpha}2, 8-Sialyltransferase from Mouse Lung1

Yukiko Yoshida, Naoya Kojima and Shuichi Tsuji2

Molecular Glycobiology, Frontier Research Program, The Institute of Physical and Chemical Research (RIKEN) Wako, Saitama 351-01

2To whom correspondence should be addressed. Phone: +81-48-462-1111 (Ext 6521), Fax: +8-48-462-4692

A cDNA encoding a new {alpha}2, 8-sialyltransferase (ST8Sia IV), which exhibits activity toward the {alpha}2, 3-linked sialic acids of N-linked oligosaccharides, was cloned from a mouse lung cDNA library by means of the PCR-based approach. The predicted amino acid sequence of ST8Sia IV showed 15.2, 56.0, and 26.2% identity with those of so far cloned mouse {alpha}2, 8-sialyltransferases, i.e., GD3 synthase (ST8Sia I), STX (ST8Sia II), and Sia{alpha}2, 3Galß1, 4GlcNAc {alpha}2, 8-sialyltransferase (ST8Sia III). ST8Sia IV exhibits high amino acid sequence identity (99.2%) with recently cloned hamster polysialyltransferase-1 gene, which is necessary to polysialic acid expression, but no enzymatic activity of the gene product was reported (Eckhardt, M. et al.(1995) Nature 373, 715-718). The ST8Sia IV gene was strongly expressed in lung, heart, and spleen, but only weak expression of the gene was observed in brain, without remarkable developmental regulation. The activity of mouse ST8Sia IV was specific toward sialylated glycoproteins. The linkage-specific sialidase treatment of glycoproteins as well as N-linked oligosaccharides from the glycoproteins revealed that ST8Sia IV exhibits an {alpha}2, 8-sialyltransferase activity toward {alpha}2, 3-linked sialic acids of N-linked oligosaccharides. In addition, ST8Sia IV can synthesize polysialic acid chain in vitro without any initiator sialyltransferase.

1This work was supported by the following grants: Grants-in-Aid for Scientific Research on Priority Areas, Nos. 0625213 (to S.T.), 07279249, 07273274 (to S.T.), and 05274103 (to S.T.), and for Encouragement of Young Scientists, No. 06770042 (to N.K.), from the Ministry of Education, Science and Culture of Japan. The nucleotide sequences reported in this paper have been submitted to the Gen Bank TM/EMBL Data Bank under accession number, X86000 [GenBank] .


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