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J. Biol. Chem., Vol. 281, Issue 16, 10945-10953, April 21, 2006

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Molecular Cloning and Characterization of a Novel 3'-Phosphoadenosine 5'-Phosphosulfate Transporter, PAPST2^*

Shin Kamiyama, Norihiko Sasaki^¶, Emi Goda^¶, Kumiko Ui-Tei^||, Kaoru Saigo^||, Hisashi Narimatsu^**, Yoshifumi Jigami^**, Reiji Kannagi, Tatsuro Irimura, and Shoko Nishihara^¶1

From the Laboratory of Cell Biology, Department of Bioinformatics, Soka University, 1-236 Tangi-cho, Hachioji, Tokyo 192-8577, Research Association for Biotechnology, 3-9, Nishi-Shinbashi 2-chome, Minato-ku, Tokyo 105-0003, ^¶Core Research for Evolutional Science and Technology of Japan Science and Technology Agency, Kawaguchi Center Building, 4-1-8, Hon-cho, Kawaguchi, Saitama 332-0012, ^||Department of Biophysics and Biochemistry, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, ^**Research Center for Glycoscience, National Institute of Advanced Industrial Science and Technology, 1-1-1 Umezono, Tsukuba, Ibaraki, 305-8586, Program of Molecular Pathology, Aichi Cancer Center, 1-1 Kanokoden, Chikusa-ku, Nagoya 464-8681, and Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan

Sulfation is an important posttranslational modification associated with a variety of molecules. It requires the involvement of the high energy form of the universal sulfate donor, 3'-phosphoadenosine 5'-phosphosulfate (PAPS). Recently, we identified a PAPS transporter gene in both humans and Drosophila. Although human colonic epithelial tissues express many sulfated glycoconjugates, PAPST1 expression in the colon is trace. In the present study, we identified a novel human PAPS transporter gene that is closely related to human PAPST1. This gene, called PAPST2, is predominantly expressed in human colon tissues. The PAPST2 protein is localized on the Golgi apparatus in a manner similar to the PAPST1 protein. By using yeast expression studies, PAPST2 protein was shown to have PAPS transport activity with an apparent K_m value of 2.2 µM, which is comparable with that of PAPST1 (0.8 µM). Overexpression of either the PAPST1 or PAPST2 gene increased PAPS transport activity in human colon cancer HCT116 cells. The RNA interference of the PAPST2 gene in the HCT116 cells significantly reduced the reactivity of G72 antibody directed against the sialyl 6-sulfo N-acetyllactosamine epitope and total sulfate incorporation into cellular proteins. These findings indicate that PAPST2 is a PAPS transporter gene involved in the synthesis of sulfated glycoconjugates in the colon.

Received for publication, August 15, 2005 , and in revised form, February 8, 2006.

The nucleotide sequence(s) reported in this paper has been submitted to the DDBJ/GenBank^TM/EBI Data Bank with accession number(s) AB231931 [GenBank] .

^* This work was supported by Core Research for Evolutional Science and Technology of the Japan Science and Technology Agency and the New Energy and Industrial Technology Development Organization. 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.

¹ To whom correspondence should be addressed. Tel./Fax: 81-426-91-8140; E-mail: shoko{at}t.soka.ac.jp.

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