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J. Biol. Chem., Vol. 278, Issue 24, 21415-21420, June 13, 2003

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Initiation of the Decorin Glycosaminoglycan Chain in the Endoplasmic Reticulum-Golgi Intermediate Compartment^*

Mats Jönsson, Erik Eklund, Lars-Åke Fransson and Åke Oldberg 

From the Department of Cell and Molecular Biology, Lund University, BMC C13, SE-221 84, Lund, Sweden

We have transiently expressed decorin with a C-terminal KDEL endoplasmic reticulum retention signal peptide in COS-7 cells to study initiation of galactosaminoglycan synthesis in the endoplasmic reticulum-Golgi intermediate compartment. All decorin-KDEL molecules were substituted with N-linked oligosaccharides sensitive to endoglycosidase H, indicating that the core protein was located proximal to the medial-Golgi. O-Linked glycosylation was only initiated in a minor fraction of the molecules. The O-linked saccharides were characterized by gel filtration after stepwise degradations using chondroitin ABC/AC-I lyases, 1–3-glycuronidase, -galactosidase, and alkaline phosphatase. The major O-linked saccharide was the linkage region pentasaccharide GalNAc1–4GlcUA1–3Gal1–3Gal1–4Xyl-2-phosphate, demonstrating initiation of chondroitin synthesis in the endoplasmic reticulum-Golgi intermediate compartment. In the presence of brefeldin A, partial elongation of a chondroitin chain took place, indicating retrieval of polymerases but not of sulfotransferases.

Received for publication, October 28, 2002 , and in revised form, March 31, 2003.

^* This work was supported by Swedish Research Council Grants 5670 and 7478, by the program "Glycoconjugates in Biological Systems" sponsored by the Swedish Foundation for Strategic Research, Greta och Johan Kock's Stiftelse, Gustav V 80-års fond, and Alfred Österlund's Stiftelse. 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.: 46-46-222-8577; Fax: 46-46-222-3128; E-mail: Ake.Oldberg{at}medkem.lu.se.

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