Structural and mechanistic characterization of bifunctional heparan sulfate N-deacetylase-N-sulfotransferase 1
Abstract Heparan sulfate (HS) polysaccharides are major constituents of the extracellular matrix, involved in myriad structural and signaling processes. Mature HS polysaccharides contain complex, non-templated patterns of sulfation and epimerization, which mediate interactions with diverse protein partners. Complex HS modifications form around initial clusters of glucosamine-N-sulfate (GlcNS) on nascent polysaccharide chains, but the mechanistic basis underpinning incorporation of the GlcNS modification itself into HS remains unclear. We have determined cryo-electron microscopy structures of human N-deacetylase-N-sulfotransferase (NDST)1, the bifunctional enzyme responsible for initial GlcNS modification of HS. Our structures reveal the architecture of both NDST1 deacetylase and sulfotransferase catalytic domains, alongside a previously unreported non-catalytic N-terminal domain. Surprisingly, the two catalytic domains of NDST1 adopt an unusual back-to-back topology that limits direct cooperativity. Binding analyses, aided by novel activity modulating nanobodies, suggest that sulfotransferase domain substrate anchoring initiates the NDST1 catalytic cycle, providing a plausible mechanism for cooperativity despite spatial domain separation. Our data shed light on key determinants of NDST1 activity, and describe tools to probe NDST1 functionin vitroandin vivo..
| Medienart: |
|
|---|
Preprint| Erscheinungsjahr: |
2023 |
|---|---|
| Erschienen: |
2023 |
| Enthalten in: |
bioRxiv.org - (2023) vom: 06. Sept. Zur Gesamtaufnahme - year:2023 |
|---|
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Mycroft-West, Courtney J. [VerfasserIn] |
|---|
| Links: |
Volltext [kostenfrei] |
|---|
| Themen: |
|---|
| doi: |
10.1101/2023.08.30.555497 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
XBI040710211 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | XBI040710211 | ||
| 003 | DE-627 | ||
| 005 | 20231205144256.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 230902s2023 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1101/2023.08.30.555497 |2 doi | |
| 035 | |a (DE-627)XBI040710211 | ||
| 035 | |a (biorXiv)10.1101/2023.08.30.555497 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Mycroft-West, Courtney J. |e verfasserin |0 (orcid)0000-0002-4963-363X |4 aut | |
| 245 | 1 | 0 | |a Structural and mechanistic characterization of bifunctional heparan sulfate N-deacetylase-N-sulfotransferase 1 |
| 264 | 1 | |c 2023 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a Computermedien |b c |2 rdamedia | ||
| 338 | |a Online-Ressource |b cr |2 rdacarrier | ||
| 520 | |a Abstract Heparan sulfate (HS) polysaccharides are major constituents of the extracellular matrix, involved in myriad structural and signaling processes. Mature HS polysaccharides contain complex, non-templated patterns of sulfation and epimerization, which mediate interactions with diverse protein partners. Complex HS modifications form around initial clusters of glucosamine-N-sulfate (GlcNS) on nascent polysaccharide chains, but the mechanistic basis underpinning incorporation of the GlcNS modification itself into HS remains unclear. We have determined cryo-electron microscopy structures of human N-deacetylase-N-sulfotransferase (NDST)1, the bifunctional enzyme responsible for initial GlcNS modification of HS. Our structures reveal the architecture of both NDST1 deacetylase and sulfotransferase catalytic domains, alongside a previously unreported non-catalytic N-terminal domain. Surprisingly, the two catalytic domains of NDST1 adopt an unusual back-to-back topology that limits direct cooperativity. Binding analyses, aided by novel activity modulating nanobodies, suggest that sulfotransferase domain substrate anchoring initiates the NDST1 catalytic cycle, providing a plausible mechanism for cooperativity despite spatial domain separation. Our data shed light on key determinants of NDST1 activity, and describe tools to probe NDST1 functionin vitroandin vivo. | ||
| 650 | 4 | |a Biology |7 (dpeaa)DE-84 | |
| 650 | 4 | |a 570 |7 (dpeaa)DE-84 | |
| 700 | 1 | |a Abdelkarim, Sahar |4 aut | |
| 700 | 1 | |a Duyvesteyn, Helen M. E. |0 (orcid)0000-0002-4641-5442 |4 aut | |
| 700 | 1 | |a Gandhi, Neha S. |0 (orcid)0000-0003-3119-6731 |4 aut | |
| 700 | 1 | |a Skidmore, Mark A. |0 (orcid)0000-0002-0287-5594 |4 aut | |
| 700 | 1 | |a Owens, Raymond J. |0 (orcid)0000-0002-3705-2993 |4 aut | |
| 700 | 1 | |a Wu, Liang |0 (orcid)0000-0003-0294-7065 |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t bioRxiv.org |g (2023) vom: 06. Sept. |
| 773 | 1 | 8 | |g year:2023 |g day:06 |g month:09 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1101/2023.08.30.555497 |z kostenfrei |3 Volltext |
| 912 | |a GBV_XBI | ||
| 951 | |a AR | ||
| 952 | |j 2023 |b 06 |c 09 | ||