Oligomerization of hamster UDP-GlcNAc:dolichol-P GlcNAc-1-P transferase, an enzyme with multiple transmembrane spans.

The Journal of Biological Chemistry
N Dan, M A Lehrman

Abstract

Hamster UDP-GlcNAc:dolichol-P GlcNAc-1-P transferase (GPT), which initiates N-linked glycosylation by catalyzing the synthesis of GlcNAc-P-P-dolichol, has multiple transmembrane spans and a catalytic site that probably exists on the cytosolic face of the endoplasmic reticulum membrane (Dan, N., Middleton, R. M., and Lehrman, M. A. (1996) J. Biol. Chem. 271, 30717-30725). In this report, we demonstrate that GPT forms functional oligomers, probably dimers. Oligomers were detected by chemical cross-linking of GPT and by a dominant-negative effect caused by co-expression of enzymatically inactive (but properly folded) GPT mutants. The GPT mutants had no effect on two other dolichol-P-dependent endoplasmic reticulum enzymes. Mixing experiments indicated that mature GPT was competent for oligomerization. Oligomerization appeared to be favored in detergent extracts compared with intact microsomes. Detergent treatments were found to prevent, rather than promote, nonspecific aggregation of GPT. These results demonstrate that GPT subunits can physically interact and influence each other. The implications of oligomerization for enzyme function are discussed. From these results, we conclude that GPT is one of a very small number of multitr...Continue Reading

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Citations

Apr 13, 2000·Proceedings of the National Academy of Sciences of the United States of America·T G BernhardtR Young
Mar 20, 1999·The Journal of Biological Chemistry·S Strahl-Bolsinger, A Scheinost
Dec 29, 1998·The Journal of Biological Chemistry·D L MarksR E Pagano
Apr 15, 2000·The Journal of Biological Chemistry·V GirrbachS Strahl-Bolsinger
Oct 6, 2000·The Journal of Biological Chemistry·J LiW W Young
Jun 8, 1999·The Journal of Biological Chemistry·H H ChengR D Simoni
Feb 21, 2018·Nature Structural & Molecular Biology·Jiho YooSeok-Yong Lee
Nov 26, 1999·Biochimica Et Biophysica Acta·M A FergusonT K Smith

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