Structural and biochemical insight into a modular β-1,4-galactan synthase in plants.


Journal

Nature plants
ISSN: 2055-0278
Titre abrégé: Nat Plants
Pays: England
ID NLM: 101651677

Informations de publication

Date de publication:
03 2023
Historique:
received: 15 07 2022
accepted: 25 01 2023
pubmed: 1 3 2023
medline: 23 3 2023
entrez: 28 2 2023
Statut: ppublish

Résumé

Rhamnogalacturonan I (RGI) is a structurally complex pectic polysaccharide with a backbone of alternating rhamnose and galacturonic acid residues substituted with arabinan and galactan side chains. Galactan synthase 1 (GalS1) transfers galactose and arabinose to either extend or cap the β-1,4-galactan side chains of RGI, respectively. Here we report the structure of GalS1 from Populus trichocarpa, showing a modular protein consisting of an N-terminal domain that represents the founding member of a new family of carbohydrate-binding module, CBM95, and a C-terminal glycosyltransferase family 92 (GT92) catalytic domain that adopts a GT-A fold. GalS1 exists as a dimer in vitro, with stem domains interacting across the chains in a 'handshake' orientation that is essential for maintaining stability and activity. In addition to understanding the enzymatic mechanism of GalS1, we gained insight into the donor and acceptor substrate binding sites using deep evolutionary analysis, molecular simulations and biochemical studies. Combining all the results, a mechanism for GalS1 catalysis and a new model for pectic galactan side-chain addition are proposed.

Identifiants

pubmed: 36849618
doi: 10.1038/s41477-023-01358-4
pii: 10.1038/s41477-023-01358-4
pmc: PMC10115243
mid: NIHMS1881134
doi:

Substances chimiques

Galactans 0
Glycosyltransferases EC 2.4.-

Types de publication

Journal Article Research Support, Non-U.S. Gov't Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S.

Langues

eng

Sous-ensembles de citation

IM

Pagination

486-500

Subventions

Organisme : NIGMS NIH HHS
ID : P30 GM124169
Pays : United States
Organisme : NIGMS NIH HHS
ID : P41 GM103390
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM130915
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM139656
Pays : United States
Organisme : US Department of Energy
ID : DE-AC02-05CH11231
Organisme : NIGMS NIH HHS
ID : P30 GM124169
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM139656
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM130915
Pays : United States

Informations de copyright

© 2023. The Author(s), under exclusive licence to Springer Nature Limited.

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Auteurs

Pradeep Kumar Prabhakar (PK)

Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, USA.
Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA.
Center for Bioenergy Innovation, Oak Ridge National Laboratory, Oakridge, TN, USA.

Jose Henrique Pereira (JH)

Molecular Biophysics and Integrated Bioimaging, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
Joint BioEnergy Institute, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.

Rahil Taujale (R)

Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, USA.

Wanchen Shao (W)

Joint BioEnergy Institute, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.

Vivek S Bharadwaj (VS)

Renewable Resources and Enabling Sciences Center, National Renewable Energy Laboratory, Golden, CO, USA.

Digantkumar Chapla (D)

Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA.

Jeong-Yeh Yang (JY)

Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA.

Yannick J Bomble (YJ)

Biosciences Center, National Renewable Energy Laboratory, Golden, CO, USA.

Kelley W Moremen (KW)

Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, USA.
Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA.

Natarajan Kannan (N)

Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, USA.

Michal Hammel (M)

Molecular Biophysics and Integrated Bioimaging, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.

Paul D Adams (PD)

Molecular Biophysics and Integrated Bioimaging, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
Joint BioEnergy Institute, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
Department of Bioengineering, University of California, Berkeley, CA, USA.

Henrik V Scheller (HV)

Joint BioEnergy Institute, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
Environmental Genomics and Systems Biology Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA.
Department of Plant and Microbial Biology, University of California, Berkeley, CA, USA.

Breeanna R Urbanowicz (BR)

Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, USA. breeanna@uga.edu.
Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA. breeanna@uga.edu.
Center for Bioenergy Innovation, Oak Ridge National Laboratory, Oakridge, TN, USA. breeanna@uga.edu.

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