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Title: Cloning, expression, crystallization and preliminary X-ray analysis of the XMT and DXMT N-methyltransferases from Coffea canephora (robusta)

Abstract

The genes encoding XMT and DXMT, the enzymes from Coffea canephora (robusta) that catalyse the three independent N-methyl transfer reactions in the caffeine-biosynthesis pathway, have been cloned and the proteins have been expressed in Escherichia coli. Both proteins have been crystallized in the presence of the demethylated cofactor S-adenosyl-l-cysteine (SAH) and substrate (xanthosine for XMT and theobromine for DXMT). Caffeine is a secondary metabolite produced by a variety of plants including Coffea canephora (robusta) and there is growing evidence that caffeine is part of a chemical defence strategy protecting young leaves and seeds from potential predators. The genes encoding XMT and DXMT, the enzymes from Coffea canephora (robusta) that catalyse the three independent N-methyl transfer reactions in the caffeine-biosynthesis pathway, have been cloned and the proteins have been expressed in Escherichia coli. Both proteins have been crystallized in the presence of the demethylated cofactor S-adenosyl-l-cysteine (SAH) and substrate (xanthosine for XMT and theobromine for DXMT). The crystals are orthorhombic, with space group P2{sub 1}2{sub 1}2{sub 1} for XMT and C222{sub 1} for DXMT. X-ray diffraction to 2.8 Å for XMT and to 2.5 Å for DXMT have been collected on beamline ID23-1 at the ESRF.

Authors:
 [1];  [2];  [3];  [2];  [3];  [2];  [4]
  1. European Molecular Biology Laboratory, 6 Rue Jules Horowitz, BP 181, 38042 Grenoble (France)
  2. Nestlé Research and Development, 101 Avenue Gustave Eiffel, Notre-Dame D’Oe, 37097 Tours (France)
  3. Department of Plant Breeding and Genetics, Department of Plant Biology, Cornell University, Ithaca, NY 14853 (United States)
  4. (France)
Publication Date:
OSTI Identifier:
22360299
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F; Journal Volume: 63; Journal Issue: Pt 4; Other Information: PMCID: PMC2330209; PMID: 17401201; PUBLISHER-ID: gj5015; OAI: oai:pubmedcentral.nih.gov:2330209; Copyright (c) International Union of Crystallography 2007; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United Kingdom
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALS; CYSTEINE; ESCHERICHIA COLI; EUROPEAN SYNCHROTRON RADIATION FACILITY; POTENTIALS; SPACE GROUPS; SUBSTRATES; X-RAY DIFFRACTION

Citation Formats

McCarthy, Andrew A., E-mail: andrewmc@embl.fr, Biget, Laurent, Lin, Chenwei, Petiard, Vincent, Tanksley, Steve D., McCarthy, James G., and European Molecular Biology Laboratory, 6 Rue Jules Horowitz, BP 181, 38042 Grenoble. Cloning, expression, crystallization and preliminary X-ray analysis of the XMT and DXMT N-methyltransferases from Coffea canephora (robusta). United Kingdom: N. p., 2007. Web. doi:10.1107/S1744309107009268.
McCarthy, Andrew A., E-mail: andrewmc@embl.fr, Biget, Laurent, Lin, Chenwei, Petiard, Vincent, Tanksley, Steve D., McCarthy, James G., & European Molecular Biology Laboratory, 6 Rue Jules Horowitz, BP 181, 38042 Grenoble. Cloning, expression, crystallization and preliminary X-ray analysis of the XMT and DXMT N-methyltransferases from Coffea canephora (robusta). United Kingdom. doi:10.1107/S1744309107009268.
McCarthy, Andrew A., E-mail: andrewmc@embl.fr, Biget, Laurent, Lin, Chenwei, Petiard, Vincent, Tanksley, Steve D., McCarthy, James G., and European Molecular Biology Laboratory, 6 Rue Jules Horowitz, BP 181, 38042 Grenoble. Sun . "Cloning, expression, crystallization and preliminary X-ray analysis of the XMT and DXMT N-methyltransferases from Coffea canephora (robusta)". United Kingdom. doi:10.1107/S1744309107009268.
@article{osti_22360299,
title = {Cloning, expression, crystallization and preliminary X-ray analysis of the XMT and DXMT N-methyltransferases from Coffea canephora (robusta)},
author = {McCarthy, Andrew A., E-mail: andrewmc@embl.fr and Biget, Laurent and Lin, Chenwei and Petiard, Vincent and Tanksley, Steve D. and McCarthy, James G. and European Molecular Biology Laboratory, 6 Rue Jules Horowitz, BP 181, 38042 Grenoble},
abstractNote = {The genes encoding XMT and DXMT, the enzymes from Coffea canephora (robusta) that catalyse the three independent N-methyl transfer reactions in the caffeine-biosynthesis pathway, have been cloned and the proteins have been expressed in Escherichia coli. Both proteins have been crystallized in the presence of the demethylated cofactor S-adenosyl-l-cysteine (SAH) and substrate (xanthosine for XMT and theobromine for DXMT). Caffeine is a secondary metabolite produced by a variety of plants including Coffea canephora (robusta) and there is growing evidence that caffeine is part of a chemical defence strategy protecting young leaves and seeds from potential predators. The genes encoding XMT and DXMT, the enzymes from Coffea canephora (robusta) that catalyse the three independent N-methyl transfer reactions in the caffeine-biosynthesis pathway, have been cloned and the proteins have been expressed in Escherichia coli. Both proteins have been crystallized in the presence of the demethylated cofactor S-adenosyl-l-cysteine (SAH) and substrate (xanthosine for XMT and theobromine for DXMT). The crystals are orthorhombic, with space group P2{sub 1}2{sub 1}2{sub 1} for XMT and C222{sub 1} for DXMT. X-ray diffraction to 2.8 Å for XMT and to 2.5 Å for DXMT have been collected on beamline ID23-1 at the ESRF.},
doi = {10.1107/S1744309107009268},
journal = {Acta Crystallographica. Section F},
number = Pt 4,
volume = 63,
place = {United Kingdom},
year = {Sun Apr 01 00:00:00 EDT 2007},
month = {Sun Apr 01 00:00:00 EDT 2007}
}
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