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Article
Nature Structural & Molecular Biology  11, 128 - 134 (2004)
Published online: 4 January 2004; | doi:10.1038/nsmb715

The ER protein folding sensor UDP-glucose glycoprotein−glucosyltransferase modifies substrates distant to local changes in glycoprotein conformation

Sean C Taylor1, 2, 3, 4, Andrew D Ferguson3, 4, John J M Bergeron2 & David Y Thomas1, 2

1  Biochemistry Department, Faculty of Medicine, McGill University, McIntyre Medical Sciences Building, 3655 Boulevard Sir William Osler, Montreal, Quebec, Canada, H3G 1Y6.

2  Department of Anatomy and Cell Biology, McGill University, Montreal, Quebec, Canada, H3A 2B2.

3  Present addresses: Montreal Proteomics Network, Genome Quebec Innovation Centre, 740 Dr. Penfield, McGill University, Montreal, Quebec, Canada H3A 2B2 (S.C.T.) and Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9050, USA (A.D.F.).

4  These authors contributed equally to this work.

Correspondence should be addressed to David Y Thomas david.thomas@mcgill.ca
We present in vitro data that explain the recognition mechanism of misfolded glycoproteins by UDP-glucose glycoprotein−glucosyltransferase (UGGT). The glycoprotein exo-(1,3)-beta-glucanase (beta-Glc) bearing two glycans unfolds in a pH-dependent manner to become a misfolded substrate for UGGT. In the crystal structure of this glycoprotein, the local hydrophobicity surrounding each glycosylation site coincides with the differential recognition of N-linked glycans by UGGT. We introduced a single F280S point mutation, producing a beta-Glc protein with full enzymatic activity that was both recognized as misfolded and monoglucosylated by UGGT. Contrary to current views, these data show that UGGT can modify N-linked glycans positioned at least 40 Å from localized regions of disorder and sense subtle conformational changes within structurally compact, enzymatically active glycoprotein substrates.

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Nature Structural & Molecular Biology
ISSN: 1545-9993
EISSN: 1545-9985
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