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Title: Structures of NodZ [alpha]1,6-fucosyltransferase in complex with GDP and GDP-fucose

Abstract

Rhizobial NodZ {alpha}1,6-fucosyltransferase ({alpha}1,6-FucT) catalyzes the transfer of the fucose (Fuc) moiety from guanosine 5'-diphosphate-{beta}-L-fucose to the reducing end of the chitin oligosaccharide core during Nod-factor (NF) biosynthesis. NF is a key signaling molecule required for successful symbiosis with a legume host for atmospheric nitrogen fixation. To date, only two {alpha}1,6-FucT structures have been determined, both without any donor or acceptor molecule that could highlight the structural background of the catalytic mechanism. Here, the first crystal structures of {alpha}1,6-FucT in complex with its substrate GDP-Fuc and with GDP, which is a byproduct of the enzymatic reaction, are presented. The crystal of the complex with GDP-Fuc was obtained through soaking of native NodZ crystals with the ligand and its structure has been determined at 2.35 {angstrom} resolution. The fucose residue is exposed to solvent and is disordered. The enzyme-product complex crystal was obtained by cocrystallization with GDP and an acceptor molecule, penta-N-acetyl-L-glucosamine (penta-NAG). The structure has been determined at 1.98 {angstrom} resolution, showing that only the GDP molecule is present in the complex. In both structures the ligands are located in a cleft formed between the two domains of NodZ and extend towards the C-terminal domain, but their conformations differ significantly.more » The structures revealed that residues in three regions of the C-terminal domain, which are conserved among {alpha}1,2-, {alpha}1,6- and protein O-fucosyltransferases, are involved in interactions with the sugar-donor molecule. There is also an interaction with the side chain of Tyr45 in the N-terminal domain, which is very unusual for a GT-B-type glycosyltransferase. Only minor conformational changes of the protein backbone are observed upon ligand binding. The only exception is a movement of the loop located between strand {beta}C2 and helix {alpha}C3. In addition, there is a shift of the {alpha}C3 helix itself upon GDP-Fuc binding.« less

Authors:
; ;  [1]
  1. NCI
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
FOREIGNNIHNCI
OSTI Identifier:
1034200
Resource Type:
Journal Article
Journal Name:
Acta Crystallogr. D
Additional Journal Information:
Journal Volume: 68; Journal Issue: (2) ; 02, 2012; Journal ID: ISSN 0907-4449
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; BIOSYNTHESIS; CHITIN; CONFORMATIONAL CHANGES; CRYSTAL STRUCTURE; GUANOSINE; HEXOSES; NITROGEN FIXATION; OLIGOSACCHARIDES; PROTEINS; RESIDUES; RESOLUTION; SOLVENTS; SUBSTRATES; SYMBIOSIS

Citation Formats

Brzezinski, Krzysztof, Dauter, Zbigniew, Jaskolski, Mariusz, and Polish). Structures of NodZ [alpha]1,6-fucosyltransferase in complex with GDP and GDP-fucose. United States: N. p., 2012. Web. doi:10.1107/S0907444911053157.
Brzezinski, Krzysztof, Dauter, Zbigniew, Jaskolski, Mariusz, & Polish). Structures of NodZ [alpha]1,6-fucosyltransferase in complex with GDP and GDP-fucose. United States. https://doi.org/10.1107/S0907444911053157
Brzezinski, Krzysztof, Dauter, Zbigniew, Jaskolski, Mariusz, and Polish). 2012. "Structures of NodZ [alpha]1,6-fucosyltransferase in complex with GDP and GDP-fucose". United States. https://doi.org/10.1107/S0907444911053157.
@article{osti_1034200,
title = {Structures of NodZ [alpha]1,6-fucosyltransferase in complex with GDP and GDP-fucose},
author = {Brzezinski, Krzysztof and Dauter, Zbigniew and Jaskolski, Mariusz and Polish)},
abstractNote = {Rhizobial NodZ {alpha}1,6-fucosyltransferase ({alpha}1,6-FucT) catalyzes the transfer of the fucose (Fuc) moiety from guanosine 5'-diphosphate-{beta}-L-fucose to the reducing end of the chitin oligosaccharide core during Nod-factor (NF) biosynthesis. NF is a key signaling molecule required for successful symbiosis with a legume host for atmospheric nitrogen fixation. To date, only two {alpha}1,6-FucT structures have been determined, both without any donor or acceptor molecule that could highlight the structural background of the catalytic mechanism. Here, the first crystal structures of {alpha}1,6-FucT in complex with its substrate GDP-Fuc and with GDP, which is a byproduct of the enzymatic reaction, are presented. The crystal of the complex with GDP-Fuc was obtained through soaking of native NodZ crystals with the ligand and its structure has been determined at 2.35 {angstrom} resolution. The fucose residue is exposed to solvent and is disordered. The enzyme-product complex crystal was obtained by cocrystallization with GDP and an acceptor molecule, penta-N-acetyl-L-glucosamine (penta-NAG). The structure has been determined at 1.98 {angstrom} resolution, showing that only the GDP molecule is present in the complex. In both structures the ligands are located in a cleft formed between the two domains of NodZ and extend towards the C-terminal domain, but their conformations differ significantly. The structures revealed that residues in three regions of the C-terminal domain, which are conserved among {alpha}1,2-, {alpha}1,6- and protein O-fucosyltransferases, are involved in interactions with the sugar-donor molecule. There is also an interaction with the side chain of Tyr45 in the N-terminal domain, which is very unusual for a GT-B-type glycosyltransferase. Only minor conformational changes of the protein backbone are observed upon ligand binding. The only exception is a movement of the loop located between strand {beta}C2 and helix {alpha}C3. In addition, there is a shift of the {alpha}C3 helix itself upon GDP-Fuc binding.},
doi = {10.1107/S0907444911053157},
url = {https://www.osti.gov/biblio/1034200}, journal = {Acta Crystallogr. D},
issn = {0907-4449},
number = (2) ; 02, 2012,
volume = 68,
place = {United States},
year = {Mon Mar 26 00:00:00 EDT 2012},
month = {Mon Mar 26 00:00:00 EDT 2012}
}