Molecular cloning and characterization of a novel 3'-phosphoadenosine 5'-phosphosulfate transporter, PAPST2

doi:10.1074/jbc.M508991200

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Papers In Press, published online ahead of print February 21, 2006
J. Biol. Chem, 10.1074/jbc.M508991200
Submitted on August 15, 2005
Revised on February 8, 2006
Accepted on February 21, 2006

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

Laboratory of Cell Biology, Department of Bioinformatics, Faculty of Engineering,, Soka University, Hachioji, Tokyo 192-8577

Corresponding Author: shoko{at}t.soka.ac.jp

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 the 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 $mu$ M, which is comparable with that of PAPST1 (0.8 $mu$ M). 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.

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