Printz B.,Luxembourg Institute of Science and Technology |
Printz B.,Universitei Catholique Of Leuven |
Dos Santos Morais R.,Luxembourg Institute of Science and Technology |
Wienkoop S.,University of Vienna |
And 4 more authors.
Frontiers in Plant Science | Year: 2015
Cell wall proteins were extracted from alfalfa stems according to a three-steps extraction procedure using sequentially CaCl2, EGTA, and LiCl-complemented buffers. The efficiency of this protocol for extracting cell wall proteins was compared with the two previously published methods optimized for alfalfa stem cell wall protein analysis. Following LC-MS/MS analysis the three-steps extraction procedure resulted in the identification of the highest number of cell wall proteins (242 NCBInr identifiers) and gave the lowest percentage of non-cell wall proteins (about 30%). However, the three protocols are rather complementary than substitutive since 43% of the identified proteins were specific to one protocol. This three-step protocol was therefore selected for a more detailed proteomic characterization using 2D-gel electrophoresis. With this technique, 75% of the identified proteins were shown to be fraction-specific and 72.7% were predicted as belonging to the cell wall compartment. Although, being less sensitive than LC-MS/MS approaches in detecting and identifying low-abundant proteins, gel-based approaches are valuable tools for the differentiation and relative quantification of protein isoforms and/or modified proteins. In particular isoforms, having variations in their amino-acid sequence and/or carrying different N-linked glycan chains were detected and characterized. This study highlights how the extracting protocols as well as the analytical techniques devoted to the study of the plant cell wall proteome are complementary and how they may be combined to elucidate the dynamism of the plant cell wall proteome in biological studies. Data are available via ProteomeXchange with identifier PXD001927. © 2015 Printz, Dos Santos Morais, Wienkoop, Sergeant, Lutts, Hausman and Renaut.
Mugemana C.,Universitei Catholique Of Leuven |
Gohy J.-F.,Universitei Catholique Of Leuven |
Fustin C.-A.,Universitei Catholique Of Leuven
Langmuir | Year: 2012
A polystyrene-[Ni 2+]-poly(ethylene oxide) metallo-supramolecular block copolymer (PS-[Ni 2+]-PEO), where -[ is a terpyridine, is used to create nanoporous thin films with free terpyridine ligands homogenously distributed on the pore walls. The PS-[Ni 2+]-PEO block copolymer is synthesized by a two step assembly process, and is then self-assembled into a thin film in order to obtain PEO cylinders oriented perpendicularly to the film surface. The supramolecular junction is opened by exposing the film to an excess of a competing ligand, and the free PEO block is then rinsed away by a selective solvent. The presence of the terpyridines on the pore walls is evidenced by fluorescence spectroscopy after formation of a fluorescent complex with an europium salt. © 2012 American Chemical Society.
Kapanda C.N.,Universitei Catholique Of Leuven |
Masquelier J.,Universitei Catholique Of Leuven |
Labar G.,Universitei Catholique Of Leuven |
Muccioli G.G.,Universitei Catholique Of Leuven |
And 2 more authors.
Journal of Medicinal Chemistry | Year: 2012
Monoacylglycerol lipase (MAGL) is responsible for signal termination of 2-arachidonoylglycerol (2-AG), an endocannabinoid neurotransmitter endowed with several physiological effects. Previously, we showed that the arylthioamide scaffold represents a privileged template for designing MAGL inhibitors. A series of 37 compounds resulting from pharmacomodulations around the arylthioamide template were synthesized and tested to evaluate their inhibitory potential on MAGL activity as well as their selectivity over fatty acid amide hydrolase (FAAH), another endocannabinoid-hydrolyzing enzyme. We have identified 2,4-dinitroaryldithiocarbamate derivatives as a novel class of MAGL inhibitors. Among the synthesized compounds, we identified [2,4-dinitrophenyl-4-(4-tert- butylbenzyl)piperazine-1-carbodithioate] (CK37), as the most potent MAGL inhibitor within this series (IC 50 = 154 nM). We have also identified [2,4-dinitrophenyl-4-benzhydrylpiperazine-1-carbodithioate] (CK16) as a selective MAGL inhibitor. These compounds are irreversible MAGL inhibitors that probably act by interacting with Cys208 or Cys242 and Ser122 residues of the enzyme. Moreover, CK37 is able to raise 2-arachidonoylglycerol (2-AG) levels in intact cells. © 2012 American Chemical Society.