Parisa Missaghian, Tabea Dierker, Elham Khosrowabadi, Fredrik Axling, Inger Eriksson, Abdurrahman Ghanem, Marion Kusche-Gullberg, Sakari Kellokumpu, Lena Kjellén, A dominant negative splice variant of the heparan sulfate biosynthesis enzyme NDST1 reduces heparan sulfate sulfation, Glycobiology, Volume 32, Issue 6, June 2022, Pages 518–528, https://doi.org/10.1093/glycob/cwac004
A dominant negative splice variant of the heparan sulfate biosynthesis enzyme NDST1 reduces heparan sulfate sulfation
|Author:||Missaghian, Parisa1; Dierker, Tabea1; Khosrowabadi, Elham2;|
1Department of Medical Biochemistry and Microbiology, The Biomedical Center, Box 582, SE-75123 Uppsala, Sweden
2Faculty of Biochemistry and Molecular Medicine, Aapistie 7A, 90220 Oulu, Finland
3Department of Surgical Sciences, Uppsala University Hospital, SE-751 85 Uppsala, Sweden
4Department of Biomedicine, University of Bergen, N-5009 Bergen, Norway
|Online Access:||PDF Full Text (PDF, 1.4 MB)|
|Persistent link:|| http://urn.fi/urn:nbn:fi-fe2022071851764
|Publish Date:|| 2022-07-18
NDST1 (glucosaminyl N-deacetylase/N-sulfotransferase) is a key enzyme in heparan sulfate (HS) biosynthesis, where it is responsible for HS N-deacetylation and N-sulfation. In addition to the full length human enzyme of 882 amino acids, here designated NDST1A, a shorter form containing 825 amino acids (NDST1B) is synthesized after alternative splicing of the NDST1 mRNA. NDST1B is mostly expressed at a low level, but increased amounts are seen in several types of cancer where it is associated with shorter survival. In this study, we aimed at characterizing the enzymatic properties of NDST1B and its effect on HS biosynthesis. Purified recombinant NDST1B lacked both N-deacetylase and N-sulfotransferase activities. Interestingly, HEK293 cells overexpressing NDST1B synthesized HS with reduced sulfation and altered domain structure. Fluorescence resonance energy transfer-microscopy demonstrated that both NDST1A and NDST1B had the capacity to interact with the HS copolymerase subunits EXT1 and EXT2 and also to form NDST1A/NDST1B dimers. Since lysates from cells overexpressing NDST1B contained less NDST enzyme activity than control cells, we suggest that NDST1B works in a dominant negative manner, tentatively by replacing the active endogenous NDST1 in the enzyme complexes taking part in biosynthesis.
|Pages:||518 - 528|
|Type of Publication:||
A1 Journal article – refereed
|Field of Science:||
1182 Biochemistry, cell and molecular biology
This work was supported by The Swedish Cancer Society Grant number 20 1338 PjF, Stiftelsen för Proteoglykanforskning, and the Medical Faculty at Uppsala University.
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