References of "Boutry, Marc"
     in
Bookmark and Share    
Full Text
Peer Reviewed
See detailN-Glycosylation of an IgG antibody secreted by Nicotiana tabacum BY-2 cells can be modulated through co-expression of human β-1,4-galactosyltransferase
Navarre, Catherine; Smargiasso, Nicolas ULiege; Duvivier, Laurent et al

in Transgenic Research (2017), 26(3), 375-384

Nicotiana tabacum BY-2 suspension cells have several advantages that make them suitable for the production of full-size monoclonal antibodies which can be purified directly from the culture medium ... [more ▼]

Nicotiana tabacum BY-2 suspension cells have several advantages that make them suitable for the production of full-size monoclonal antibodies which can be purified directly from the culture medium. Carbohydrate characterization of an antibody (Lo-BM2) expressed in N. tabacum BY-2 cells showed that the purified Lo-BM2 displays N-glycan homogeneity with a high proportion (>70%) of the complex GnGnXF glycoform. The stable co-expression of a human β-1,4-galactosyltransferase targeted to different Golgi sub-compartments altered Lo-BM2N-glycosylation and resulted in the production of an antibody that exhibited either hybrid structures containing a low abundance of the plant epitopes (α-1,3-fucose and β-1,2-xylose), or a large amount of galactose-extended N-glycan structures. These results demonstrate the suitability of stable N-glycoengineered N. tabacum BY-2 cell lines for the production of human-like antibodies. © 2017, Springer International Publishing Switzerland. [less ▲]

Detailed reference viewed: 22 (3 ULiège)
Full Text
Peer Reviewed
See detailPhotosynthetic trichomes contain a specific Rubisco with a modified pH9 dependent activity
Laterre, Raphaëlle; Pottier, Mathieu; Remacle, Claire ULiege et al

in Plant Physiology (2017)

Detailed reference viewed: 64 (8 ULiège)
Full Text
Peer Reviewed
See detailInactivation of the β(1,2)-xylosyltransferase and the α(1,3)-fucosyltransferase genes in Nicotiana tabacum BY-2 cells by a multiplex CRISPR/Cas9 strategy results in glycoproteins without plant-specific glycans
Mercx, Sébastien; Smargiasso, Nicolas ULiege; Chaumont, François et al

in Frontiers in Plant Science (2017), 8

Plants or plant cells can be used to produce pharmacological glycoproteins such as antibodies or vaccines. However these proteins carry N-glycans with planttypical residues [β(1,2)-xylose and core α(1,3 ... [more ▼]

Plants or plant cells can be used to produce pharmacological glycoproteins such as antibodies or vaccines. However these proteins carry N-glycans with planttypical residues [β(1,2)-xylose and core α(1,3)-fucose], which can greatly impact the immunogenicity, allergenicity, or activity of the protein. Two enzymes are responsible for the addition of plant-specific glycans: β(1,2)-xylosyltransferase (XylT) and α(1,3)- fucosyltransferase (FucT). Our aim consisted of knocking-out two XylT genes and four FucT genes (12 alleles altogether) in Nicotiana tabacum BY-2 suspension cells using CRISPR/Cas9. Three XylT and six FucT sgRNAs were designed to target conserved regions. After transformation of N. tabacum BY-2 cells with genes coding for sgRNAs, Cas9, and a selectable marker (bar), transgenic lines were obtained and their extracellular as well as intracellular protein complements were analyzed by Western blotting using antibodies recognizing β(1,2)-xylose and α(1,3)-fucose. Three lines showed a strong reduction of β(1,2)-xylose and α(1,3)-fucose, while two lines were completely devoid of them, indicating complete gene inactivation. The absence of these carbohydrates was confirmed by mass spectrometry analysis of the extracellular proteins. PCR amplification and sequencing of the targeted region indicated small INDEL and/or deletions between the target sites. The KO lines did not show any particular morphology and grew as the wild-type. One KO line was transformed with genes encoding a human IgG2 antibody. The IgG2 expression level was as high as in a control transformant which had not been glycoengineered. The IgG glycosylation profile determined by mass spectrometry confirmed that no β(1,2)-xylose or α(1,3)-fucose were present on the glycosylation moiety and that the dominant glycoform was the GnGn structure. These data represent an important step toward humanizing the glycosylation of pharmacological proteins expressed in N. tabacum BY-2 cells. © 2017 Mercx, Smargiasso, Chaumont, De Pauw, Boutry and Navarre. [less ▲]

Detailed reference viewed: 15 (2 ULiège)
Full Text
Peer Reviewed
See detailAtypical subunit composition of the chlorophycean mitochondrial F1FO ATP synthase and role of Asa7 protein in stability and oligomycin resistance of the enzyme.
Lapaille, Marie; Escobar-Ramirez, Adelma; Degand, Hervé et al

in Molecular Biology and Evolution (2010), 27(7), 1630-1644

Background. In yeast, mammals, and land plants, mitochondrial F(1)F(O) ATP synthase (complex V) is a remarkable enzymatic machinery which comprises about 15 conserved subunits. Peculiar among eukaryotes ... [more ▼]

Background. In yeast, mammals, and land plants, mitochondrial F(1)F(O) ATP synthase (complex V) is a remarkable enzymatic machinery which comprises about 15 conserved subunits. Peculiar among eukaryotes, complex V from Chlamydomonadales algae (order of chlorophycean class) has an atypical subunit composition of its peripheral stator and dimerization module, with 9 subunits of unknown evolutionary origin (Asa subunits). In vitro, this enzyme exhibits an increased stability of its dimeric form, and in vivo, Chlamydomonas reinhardtii cells are insensitive to oligomycins, which are potent inhibitors of proton translocation through the F(O) moiety. Methodology/Principal Findings. In this work, we showed that the atypical features of the Chlamydomonadales complex V enzyme are shared by the other chlorophycean orders. By biochemical and in silico analyses, we detected several atypical Asa subunits in Scenedesmus obliquus (Sphaeropleales) and Chlorococcum ellipsoideum (Chlorococcales). In contrast, Complex V has a canonical subunit composition in other classes of Chlorophytes (Trebouxiophyceae, Prasinophyceae, and Ulvophyceae) as well as in Streptophytes (land plants) and in Rhodophytes (red algae). Growth, respiration and ATP levels in Chlorophyceae were also barely affected by oligomycin concentrations that affect representatives of the other classes of Chlorophytes. We finally studied the function of the Asa7 atypical subunit by using RNA interference in C. reinhardtii. Although the loss of Asa7 subunit has no impact on cell bioenergetics or mitochondrial structures, it destabilizes in vitro the enzyme dimeric form and renders growth, respiration and ATP level sensitive to oligomycins. Conclusions/Significance. Altogether, our results suggest that the loss of canonical components of the Complex V stator happened at the root of chlorophycean lineage and was accompanied by the recruitment of novel polypeptides. Such a massive modification of Complex V stator features might have conferred novel properties, including the stabilization of the enzyme dimeric form and the shielding of the proton channel. In these respects, we discuss an evolutionary scenario for F(1)F(O) ATP synthase in the whole green lineage (i.e. Chlorophyta and Streptophyta). [less ▲]

Detailed reference viewed: 61 (19 ULiège)
Full Text
See detailF1FO ATP synthase mutants in Chlamydomonas: Stability and oligomycin resistance mediated by atypical Asa7 protein; interaction between chloroplastic and mitochondrial bioenergetics
Lapaille, Marie ULiege; Escobar-Ramírez, Adelma; Degand, Hervé et al

in Biochimica et Biophysica Acta (BBA) - Bioenergetics (2010), 1797(Supplement 1), 29

Detailed reference viewed: 46 (10 ULiège)
Peer Reviewed
See detailAnalysis Of The Chromosome Sequence Of The Legume Symbiont Sinorhizobium Meliloti Strain 1021
Capela, Delphine; Barloy-Hubler, Frédérique; Gouzy, Jérôme et al

in Proceedings of the National Academy of Sciences of the United States of America (2001), 98(17),

Detailed reference viewed: 14 (2 ULiège)
Peer Reviewed
See detailThe Composite Genome Of The Legume Symbiont Sinorhizobium Meliloti
Galibert, Francis; Finan, Turlough M.; Long, Sharon R. et al

in Science (2001), 293(5530),

Detailed reference viewed: 31 (4 ULiège)