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J. Biol. Chem., Vol. 280, Issue 33, 29596-29603, August 19, 2005

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A Proteoglycan Undergoes Different Modifications en Route to the Apical and Basolateral Surfaces of Madin-Darby Canine Kidney Cells^*

Heidi Tveit, Gunnar Dick, Venke Skibeli, and Kristian Prydz

From the Department of Molecular Biosciences, University of Oslo, Box 1041 Blindern, 0316 Oslo, Norway

We have grown polarized epithelial Madin-Darby canine kidney II (MDCK II) cells on filters in the presence of [³⁵S]sulfate, [³H]glucosamine, or [³⁵S]cysteine/[³⁵S]methionine to study proteoglycan (PG) synthesis, sorting, and secretion to the apical and basolateral media. Whereas most of the [³⁵S]sulfate label was recovered in basolateral PGs, the [³H]glucosamine label was predominantly incorporated into the glycosaminoglycan chains of apical PGs, indicating that basolateral PGs are more intensely sulfated than their apical counterparts. Expression of the PG serglycin with a green fluorescent protein tag (SG-GFP) in MDCK II cells produced a protein core secreted 85% apically, which was largely modified by chondroitin sulfate chains. Surprisingly, the 15% of secreted SG-GFP molecules recovered basolaterally were more heavily sulfated and displayed a different sulfation pattern than the apical counterpart. More detailed studies of the differential modification of apically and basolaterally secreted SG-GFP indicate that the protein cores have been designated to apical and basolateral transport platforms before pathway-specific, post-translational modifications have been completed.

Received for publication, April 5, 2005 , and in revised form, June 20, 2005.

^* This work was supported by The Research Council of Norway, the Norwegian Cancer Society, and The Blix Foundation. 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.

To whom correspondence should be addressed. Tel.: 47-2-285-6753; Fax: 47-2-285-4443; E-mail: kristian.prydz{at}imbv.uio.no.

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