Role of UDP-N-Acetylglucosamine (GlcNAc) and O-GlcNAcylation of Hyaluronan Synthase 2 in the Control of Chondroitin Sulfate and Hyaluronan Synthesis
| Autor: | Davide Vigetti, Paola Moretto, Barbara Bartolini, Alberto Passi, Markku Tammi, Eugenia Karousou, Manuela Viola, Giancarlo De Luca, Sara Deleonibus, Vincent C. Hascall |
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| Rok vydání: | 2012 |
| Předmět: |
Glycosylation
Mutation Missense Muscle Proteins Glycobiology and Extracellular Matrices Hyaluronan Synthase 2 Biochemistry Muscle Smooth Vascular Cell Line Extracellular matrix Glycosaminoglycan Serine chemistry.chemical_compound Glucosamine Hyaluronic acid Humans Chondroitin sulfate Glucuronosyltransferase Hyaluronic Acid Molecular Biology Aorta Uridine Diphosphate N-Acetylglucosamine Chondroitin Sulfates Cell Biology carbohydrates (lipids) Amino Acid Substitution chemistry lipids (amino acids peptides and proteins) Hyaluronan Synthases Diabetic Angiopathies |
| Popis: | Hyaluronan (HA) is a glycosaminoglycan present in most tissue microenvironments that can modulate many cell behaviors, including proliferation, migration, and adhesive proprieties. In contrast with other glycosaminoglycans, which are synthesized in the Golgi, HA is synthesized at the plasma membrane by one or more of the three HA synthases (HAS1-3), which use cytoplasmic UDP-glucuronic acid and UDP-N-acetylglucosamine as substrates. Previous studies revealed the importance of UDP-sugars for regulating HA synthesis. Therefore, we analyzed the effect of UDP-GlcNAc availability and protein glycosylation with O-linked N-acetylglucosamine (O-GlcNAcylation) on HA and chondroitin sulfate synthesis in primary human aortic smooth muscle cells. Glucosamine treatment, which increases UDP-GlcNAc availability and protein O-GlcNAcylation, increased synthesis of both HA and chondroitin sulfate. However, increasing O-GlcNAcylation by stimulation with O-(2-acetamido-2-deoxy-d-glucopyranosylidene)amino-N-phenylcarbamate without a concomitant increase of UDP-GlcNAc increased only HA synthesis. We found that HAS2, the main synthase in aortic smooth muscle cells, can be O-GlcNAcylated on serine 221, which strongly increased its activity and its stability (t(frac12;)5 h versus ∼17 min without O-GlcNAcylation). S221A mutation prevented HAS2 O-GlcNAcylation, which maintained the rapid turnover rate even in the presence of GlcN and increased UDP-GlcNAc. These findings could explain the elevated matrix HA observed in diabetic vessels that, in turn, could mediate cell dedifferentiation processes critical in vascular pathologies. |
| Databáze: | OpenAIRE |
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