Dermatan sulfate epimerase 1 and dermatan 4-O-sulfotransferase 1 form complexes that generate long epimerized 4-O-sulfated blocks
| Autor: | Marco Maccarana, Ulf Ellervik, Gunilla Westergren-Thorsson, Katarzyna Zielinska, Emil Tykesson, Giacomo Frati, Antti Hassinen, Anders Malmström, Sakari Kellokumpu, Martin A. Thelin |
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| Přispěvatelé: | Institute for Molecular Medicine Finland, University of Helsinki |
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Sulfotransferase N-ACETYLGALACTOSAMINYLTRANSFERASE-1 GROWTH FACTOR/SCATTER FACTOR Iduronic acid Biochemistry dermatan sulfate Dermatan sulfate 03 medical and health sciences chemistry.chemical_compound SUBSTRATE Sulfation D4ST1 glycobiology epimerization Golgi glycosaminoglycan CHONDROITIN SULFATE BIOSYNTHESIS Chondroitin sulfate CANCER-CELLS Molecular Biology chondroitin sulfate Heparin cofactor II chemistry.chemical_classification HEPARIN-COFACTOR-II 030102 biochemistry & molecular biology epimerase Cell Biology Glucuronic acid DS-epi2 DS-epi1 POLYMERIZATION 030104 developmental biology Enzyme chemistry IDURONIC ACID 1182 Biochemistry cell and molecular biology 3111 Biomedicine EHLERS-DANLOS-SYNDROME |
| Popis: | During the biosynthesis of chondroitin/dermatan sulfate (CS/DS), a variable fraction of glucuronic acid is converted to iduronic acid through the activities of two epimerases, dermatan sulfate epimerases 1 (DS-epi1) and 2 (DS-epi2). Previous in vitro studies indicated that without association with other enzymes, DS-epi1 activity produces structures that have only a few adjacent iduronic acid units. In vivo, concomitant with epimerization, dermatan 4-O-sulfotransferase 1 (D4ST1) sulfates the GalNAc adjacent to iduronic acid. This sulfation facilitates DS-epi1 activity and enables the formation of long blocks of sulfated iduronic acid-containing domains, which can be major components of CS/DS. In this report, we used recombinant enzymes to confirm the concerted action of DS-epi1 and D4ST1. Confocal microscopy revealed that these two enzymes colocalize to the Golgi, and FRET experiments indicated that they physically interact. Furthermore, FRET, immunoprecipitation, and cross-linking experiments also revealed that DS-epi1, DS-epi2, and D4ST1 form homomers and are all part of a hetero-oligomeric complex where D4ST1 directly interacts with DS-epi1, but not with DS-epi2. The cooperation of DS-epi1 with D4ST1 may therefore explain the processive mode of the formation of iduronic acid blocks. In conclusion, the iduronic acid-forming enzymes operate in complexes, similar to other enzymes active in glycosaminoglycan biosynthesis. This knowledge shed light on regulatory mechanisms controlling the biosynthesis of the structurally diverse CS/DS molecule. |
| Databáze: | OpenAIRE |
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