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  1. Article ; Online: Salmonid polysialyltransferases to generate a variety of sialic acid polymers

    Mathieu Decloquement / Marzia Tindara Venuto / Virginie Cogez / Anna Steinmetz / Céline Schulz / Cédric Lion / Maxence Noel / Vincent Rigolot / Roxana Elin Teppa / Christophe Biot / Alexander Rebl / Sebastian Peter Galuska / Anne Harduin-Lepers

    Scientific Reports, Vol 13, Iss 1, Pp 1-

    2023  Volume 14

    Abstract: Abstract The human polysialyltransferases ST8Sia II and ST8Sia IV catalyze the transfer of several Neu5Ac residues onto glycoproteins forming homopolymers with essential roles during different physiological processes. In salmonids, heterogeneous set of ... ...

    Abstract Abstract The human polysialyltransferases ST8Sia II and ST8Sia IV catalyze the transfer of several Neu5Ac residues onto glycoproteins forming homopolymers with essential roles during different physiological processes. In salmonids, heterogeneous set of sialic acids polymers have been described in ovary and on eggs cell surface and three genes st8sia4, st8sia2-r1 and st8sia2-r2 were identified that could be implicated in these heteropolymers. The three polysialyltransferases from the salmonid Coregonus maraena were cloned, recombinantly expressed in HEK293 cells and the ST8Sia IV was biochemically characterized. The MicroPlate Sialyltransferase Assay and the non-natural donor substrate CMP-SiaNAl were used to demonstrate enzyme activity and optimize polysialylation reactions. Polysialylation was also carried out with natural donor substrates CMP-Neu5Ac, CMP-Neu5Gc and CMP-Kdn in cell-free and cell-based assays and structural analyses of polysialylated products using the anti-polySia monoclonal antibody 735 and endoneuraminidase N and HPLC approaches. Our data highlighted distinct specificities of human and salmonid polysialyltransferases with notable differences in donor substrates use and the capacity of fish enzymes to generate heteropolymers. This study further suggested an evolution of the biological functions of polySia. C. maraena ST8Sia IV of particular interest to modify glycoproteins with a variety of polySia chains.
    Keywords Medicine ; R ; Science ; Q
    Subject code 540
    Language English
    Publishing date 2023-09-01T00:00:00Z
    Publisher Nature Portfolio
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  2. Article ; Online: Exploring the Potential of β-Catenin O -GlcNAcylation by Using Fluorescence-Based Engineering and Imaging

    Angelina Kasprowicz / Corentin Spriet / Christine Terryn / Vincent Rigolot / Stephan Hardiville / Matthew G. Alteen / Tony Lefebvre / Christophe Biot

    Molecules, Vol 25, Iss 4501, p

    2020  Volume 4501

    Abstract: Monitoring glycosylation changes within cells upon response to stimuli remains challenging because of the complexity of this large family of post-translational modifications (PTMs). We developed an original tool, enabling labeling and visualization of ... ...

    Abstract Monitoring glycosylation changes within cells upon response to stimuli remains challenging because of the complexity of this large family of post-translational modifications (PTMs). We developed an original tool, enabling labeling and visualization of the cell cycle key-regulator β-catenin in its O -GlcNAcylated form, based on intramolecular Förster resonance energy transfer (FRET) technology in cells. We opted for a bioorthogonal chemical reporter strategy based on the dual-labeling of β-catenin with a green fluorescent protein (GFP) for protein sequence combined with a chemically-clicked imaging probe for PTM, resulting in a fast and easy to monitor qualitative FRET assay. We validated this technology by imaging the O -GlcNAcylation status of β-catenin in HeLa cells. The changes in O -GlcNAcylation of β-catenin were varied by perturbing global cellular O -GlcNAc levels with the inhibitors of O -GlcNAc transferase (OGT) and O -GlcNAcase (OGA). Finally, we provided a flowchart demonstrating how this technology is transposable to any kind of glycosylation.
    Keywords bioorthogonal chemistry ; fluorescence ; glycosylation ; metabolic incorporation ; GFP ; β-catenin ; Organic chemistry ; QD241-441
    Subject code 500
    Language English
    Publishing date 2020-10-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  3. Article ; Online: UDP-GLYCOSYLTRANSFERASE 72E3 Plays a Role in Lignification of Secondary Cell Walls in Arabidopsis

    Fabien Baldacci-Cresp / Julien Le Roy / Brigitte Huss / Cédric Lion / Anne Créach / Corentin Spriet / Ludovic Duponchel / Christophe Biot / Marie Baucher / Simon Hawkins / Godfrey Neutelings

    International Journal of Molecular Sciences, Vol 21, Iss 6094, p

    2020  Volume 6094

    Abstract: Lignin is present in plant secondary cell walls and is among the most abundant biological polymers on Earth. In this work we investigated the potential role of the UGT72E gene family in regulating lignification in Arabidopsis . Chemical determination of ... ...

    Abstract Lignin is present in plant secondary cell walls and is among the most abundant biological polymers on Earth. In this work we investigated the potential role of the UGT72E gene family in regulating lignification in Arabidopsis . Chemical determination of floral stem lignin contents in ugt72e1 , ugt72e2, and ugt72e3 mutants revealed no significant differences compared to WT plants. In contrast, the use of a novel safranin O ratiometric imaging technique indicated a significant increase in the cell wall lignin content of both interfascicular fibers and xylem from young regions of ugt72e3 mutant floral stems. These results were globally confirmed in interfascicular fibers by Raman microspectroscopy. Subsequent investigation using a bioorthogonal triple labelling strategy suggested that the augmentation in lignification was associated with an increased capacity of mutant cell walls to incorporate H-, G-, and S-monolignol reporters. Expression analysis showed that this increase was associated with an up-regulation of LAC17 and PRX71 , which play a key role in lignin polymerization. Altogether, these results suggest that UGT72E3 can influence the kinetics of lignin deposition by regulating monolignol flow to the cell wall as well as the potential of this compartment to incorporate monomers into the growing lignin polymer.
    Keywords lignin ; UDP glycosyltransferase ; glycosylation ; monolignol incorporation ; cell wall ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
    Subject code 580
    Language English
    Publishing date 2020-08-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  4. Article ; Online: Structure‐Based Design and Synthesis of Piperidinol‐Containing Molecules as New Mycobacterium abscessus Inhibitors

    Dr. Jérôme deRuyck / Dr. Christian Dupont / Elodie Lamy / Dr. Vincent Le Moigne / Prof. Christophe Biot / Dr. Yann Guérardel / Prof. Jean‐Louis Herrmann / Dr. Mickaël Blaise / Dr. Stanislas Grassin‐Delyle / Dr. Laurent Kremer / Dr. Faustine Dubar

    ChemistryOpen, Vol 9, Iss 3, Pp 351-

    2020  Volume 365

    Abstract: Abstract Non‐tuberculous mycobacterium (NTM) infections, such as those caused by Mycobacterium abscessus, are increasing globally. Due to their intrinsic drug resistance, M. abscessus pulmonary infections are often difficult to cure using standard ... ...

    Abstract Abstract Non‐tuberculous mycobacterium (NTM) infections, such as those caused by Mycobacterium abscessus, are increasing globally. Due to their intrinsic drug resistance, M. abscessus pulmonary infections are often difficult to cure using standard chemotherapy. We previously demonstrated that a piperidinol derivative, named PIPD1, is an efficient molecule both against M. abscessus and Mycobacterium tuberculosis, the agent of tuberculosis, by targeting the mycolic acid transporter MmpL3. These results prompted us to design and synthesize a series of piperidinol derivatives and to determine the biological activity against M. abscessus. Structure‐activity relationship (SAR) studies pointed toward specific sites on the scaffold that can tolerate slight modifications. Overall, these results identified FMD‐88 as a new promising active analogue against M. abscessus. Also, we determined the pharmacokinetics properties of PIPD1 and showed that intraperitoneal administration of this compound resulted in promising serum concentration and an elimination half‐life of 3.2 hours.
    Keywords mycobacterium abscessus ; molecular modeling ; structure-activity relationship ; phenotypic screening ; piperidinol derivatives ; Chemistry ; QD1-999
    Subject code 540
    Language English
    Publishing date 2020-03-01T00:00:00Z
    Publisher Wiley-VCH
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  5. Article ; Online: GORAB scaffolds COPI at the trans-Golgi for efficient enzyme recycling and correct protein glycosylation

    Tomasz M. Witkos / Wing Lee Chan / Merja Joensuu / Manuel Rhiel / Ed Pallister / Jane Thomas-Oates / A. Paul Mould / Alex A. Mironov / Christophe Biot / Yann Guerardel / Willy Morelle / Daniel Ungar / Felix T. Wieland / Eija Jokitalo / May Tassabehji / Uwe Kornak / Martin Lowe

    Nature Communications, Vol 10, Iss 1, Pp 1-

    2019  Volume 18

    Abstract: COPI is recruited to the membrane by binding to Arf GTPases. Here the authors find that GORAB, a trans-Golgi protein, promotes COPI recruitment by forming membrane domains that also contain the COPI-interacting protein Scyl1, which is required for ... ...

    Abstract COPI is recruited to the membrane by binding to Arf GTPases. Here the authors find that GORAB, a trans-Golgi protein, promotes COPI recruitment by forming membrane domains that also contain the COPI-interacting protein Scyl1, which is required for efficient glycosylation of cargo proteins.
    Keywords Science ; Q
    Language English
    Publishing date 2019-01-01T00:00:00Z
    Publisher Nature Publishing Group
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  6. Article ; Online: GORAB scaffolds COPI at the trans-Golgi for efficient enzyme recycling and correct protein glycosylation

    Tomasz M. Witkos / Wing Lee Chan / Merja Joensuu / Manuel Rhiel / Ed Pallister / Jane Thomas-Oates / A. Paul Mould / Alex A. Mironov / Christophe Biot / Yann Guerardel / Willy Morelle / Daniel Ungar / Felix T. Wieland / Eija Jokitalo / May Tassabehji / Uwe Kornak / Martin Lowe

    Nature Communications, Vol 10, Iss 1, Pp 1-

    2019  Volume 18

    Abstract: COPI is recruited to the membrane by binding to Arf GTPases. Here the authors find that GORAB, a trans-Golgi protein, promotes COPI recruitment by forming membrane domains that also contain the COPI-interacting protein Scyl1, which is required for ... ...

    Abstract COPI is recruited to the membrane by binding to Arf GTPases. Here the authors find that GORAB, a trans-Golgi protein, promotes COPI recruitment by forming membrane domains that also contain the COPI-interacting protein Scyl1, which is required for efficient glycosylation of cargo proteins.
    Keywords Science ; Q
    Language English
    Publishing date 2019-01-01T00:00:00Z
    Publisher Nature Portfolio
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  7. Article ; Online: Novel Zebrafish Mono-α2,8-sialyltransferase (ST8Sia VIII)

    Lan-Yi Chang / Elin Teppa / Maxence Noel / Pierre-André Gilormini / Mathieu Decloquement / Cédric Lion / Christophe Biot / Anne-Marie Mir / Virginie Cogez / Philippe Delannoy / Kay Hooi Khoo / Daniel Petit / Yann Guérardel / Anne Harduin-Lepers

    International Journal of Molecular Sciences, Vol 20, Iss 3, p

    An Evolutionary Perspective of α2,8-Sialylation

    2019  Volume 622

    Abstract: The mammalian mono-α2,8-sialyltransferase ST8Sia VI has been shown to catalyze the transfer of a unique sialic acid residues onto core 1 O -glycans leading to the formation of di-sialylated O -glycosylproteins and to a lesser extent to diSia motifs onto ... ...

    Abstract The mammalian mono-α2,8-sialyltransferase ST8Sia VI has been shown to catalyze the transfer of a unique sialic acid residues onto core 1 O -glycans leading to the formation of di-sialylated O -glycosylproteins and to a lesser extent to diSia motifs onto glycolipids like GD1a. Previous studies also reported the identification of an orthologue of the ST8SIA6 gene in the zebrafish genome. Trying to get insights into the biosynthesis and function of the oligo-sialylated glycoproteins during zebrafish development, we cloned and studied this fish α2,8-sialyltransferase homologue. In situ hybridization experiments demonstrate that expression of this gene is always detectable during zebrafish development both in the central nervous system and in non-neuronal tissues. Intriguingly, using biochemical approaches and the newly developed in vitro MicroPlate Sialyltransferase Assay (MPSA), we found that the zebrafish recombinant enzyme does not synthetize diSia motifs on glycoproteins or glycolipids as the human homologue does. Using comparative genomics and molecular phylogeny approaches, we show in this work that the human ST8Sia VI orthologue has disappeared in the ray-finned fish and that the homologue described in fish correspond to a new subfamily of α2,8-sialyltransferase named ST8Sia VIII that was not maintained in Chondrichtyes and Sarcopterygii.
    Keywords mono-α2,8-sialyltransferases ; diSia motifs ; evolution ; ST8Sia ; functional genomics ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
    Subject code 572
    Language English
    Publishing date 2019-01-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  8. Article ; Online: Ferroquine, the next generation antimalarial drug, has antitumor activity

    Artem Kondratskyi / Kateryna Kondratska / Fabien Vanden Abeele / Dmitri Gordienko / Charlotte Dubois / Robert-Allain Toillon / Christian Slomianny / Sébastien Lemière / Philippe Delcourt / Etienne Dewailly / Roman Skryma / Christophe Biot / Natalia Prevarskaya

    Scientific Reports, Vol 7, Iss 1, Pp 1-

    2017  Volume 15

    Abstract: Abstract Despite the tremendous progress in medicine, cancer remains one of the most serious global health problems awaiting new effective therapies. Here we present ferroquine (FQ), the next generation antimalarial drug, as a promising candidate for ... ...

    Abstract Abstract Despite the tremendous progress in medicine, cancer remains one of the most serious global health problems awaiting new effective therapies. Here we present ferroquine (FQ), the next generation antimalarial drug, as a promising candidate for repositioning as cancer therapeutics. We report that FQ potently inhibits autophagy, perturbs lysosomal function and impairs prostate tumor growth in vivo. We demonstrate that FQ negatively regulates Akt kinase and hypoxia-inducible factor-1α (HIF-1α) and is particularly effective in starved and hypoxic conditions frequently observed in advanced solid cancers. FQ enhances the anticancer activity of several chemotherapeutics suggesting its potential application as an adjuvant to existing anticancer therapy. Alike its parent compound chloroquine (CQ), FQ accumulates within and deacidifies lysosomes. Further, FQ induces lysosomal membrane permeabilization, mitochondrial depolarization and caspase-independent cancer cell death. Overall, our work identifies ferroquine as a promising new drug with a potent anticancer activity.
    Keywords Medicine ; R ; Science ; Q
    Subject code 610
    Language English
    Publishing date 2017-11-01T00:00:00Z
    Publisher Nature Publishing Group
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  9. Article ; Online: Ferroquine, an Ingenious Antimalarial Drug –Thoughts on the Mechanism of Action

    Faustine Dubar / Jamal Khalife / Jacques Brocard / Christophe Biot / Daniel Dive

    Molecules, Vol 13, Iss 11, Pp 2900-

    2008  Volume 2907

    Abstract: Ferroquine (FQ or SR97193) is a novel antimalarial drug candidate, currently in development at Sanofi-Aventis. In contrast to conventional drugs, FQ is the first organometallic drug: a ferrocenyl group covalently flanked by a 4-aminoquinoline and a basic ...

    Abstract Ferroquine (FQ or SR97193) is a novel antimalarial drug candidate, currently in development at Sanofi-Aventis. In contrast to conventional drugs, FQ is the first organometallic drug: a ferrocenyl group covalently flanked by a 4-aminoquinoline and a basic alkylamine. FQ is able to overcome the CQ resistance problem, an important limit to the control of Plasmodium falciparum, the principal causative agent of malaria. After fifteen years of effort, it is now possible to propose a multifactorial mechanism of action of FQ by its capacity to target lipids, to inhibit the formation of hemozoin and to generate reactive oxygen species.
    Keywords Malaria ; Bioorganometallics ; Ferroquine ; Mechanism of action ; Resistance ; Organic chemistry ; QD241-441
    Language English
    Publishing date 2008-11-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  10. Article ; Online: Ciprofloxacin Derivatives Affect Parasite Cell Division and Increase the Survival of Mice Infected with Toxoplasma gondii.

    Erica S Martins-Duarte / Faustine Dubar / Philippe Lawton / Cristiane França da Silva / Maria de Nazaré C Soeiro / Wanderley de Souza / Christophe Biot / Rossiane C Vommaro

    PLoS ONE, Vol 10, Iss 5, p e

    2015  Volume 0125705

    Abstract: Toxoplasmosis, caused by the protozoan Toxoplasma gondii, is a worldwide disease whose clinical manifestations include encephalitis and congenital malformations in newborns. Previously, we described the synthesis of new ethyl-ester derivatives of the ... ...

    Abstract Toxoplasmosis, caused by the protozoan Toxoplasma gondii, is a worldwide disease whose clinical manifestations include encephalitis and congenital malformations in newborns. Previously, we described the synthesis of new ethyl-ester derivatives of the antibiotic ciprofloxacin with ~40-fold increased activity against T. gondii in vitro, compared with the original compound. Cipro derivatives are expected to target the parasite's DNA gyrase complex in the apicoplast. The activity of these compounds in vivo, as well as their mode of action, remained thus far uncharacterized. Here, we examined the activity of the Cipro derivatives in vivo, in a model of acute murine toxoplasmosis. In addition, we investigated the cellular effects T. gondii tachyzoites in vitro, by immunofluorescence and transmission electron microscopy (TEM). When compared with Cipro treatment, 7-day treatments with Cipro derivatives increased mouse survival significantly, with 13-25% of mice surviving for up to 60 days post-infection (vs. complete lethality 10 days post-infection, with Cipro treatment). Light microscopy examination early (6 and 24h) post-infection revealed that 6-h treatments with Cipro derivatives inhibited the initial event of parasite cell division inside host cells, in an irreversible manner. By TEM and immunofluorescence, the main cellular effects observed after treatment with Cipro derivatives and Cipro were cell scission inhibition--with the appearance of 'tethered' parasites--malformation of the inner membrane complex, and apicoplast enlargement and missegregation. Interestingly, tethered daughter cells resulting from Cipro derivatives, and also Cipro, treatment did not show MORN1 cap or centrocone localization. The biological activity of Cipro derivatives against C. parvum, an apicomplexan species that lacks the apicoplast, is, approximately, 50 fold lower than that in T. gondii tachyzoites, supporting that these compounds targets the apicoplast. Our results show that Cipro derivatives improved the survival of mice acutely infected with T. gondii and inhibited parasite replication early in the first cycle of infection in vitro, highlighting their therapeutic potential for the treatment of toxoplasmosis.
    Keywords Medicine ; R ; Science ; Q
    Subject code 572
    Language English
    Publishing date 2015-01-01T00:00:00Z
    Publisher Public Library of Science (PLoS)
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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