References of "De Pauw, Edwin"
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See detailDe novo sequencing of unusual non tryptic peptides thanks to 4-sulfophenylisothiocyanate derivatization by post-source decay MALDI-MS.
Echterbille, Julien ULg; Quinton, Loïc ULg; Escoubas, Pierre et al

Poster (2013, June 11)

Introduction Due to the specificity of trypsin, tryptic peptides contain basic residues on the C-terminal side. This feature provides good ionization efficiency, and facilitates fragmentation processes ... [more ▼]

Introduction Due to the specificity of trypsin, tryptic peptides contain basic residues on the C-terminal side. This feature provides good ionization efficiency, and facilitates fragmentation processes. In the case of non tryptic peptides, the absence of basic residues at one extremity implicates lower fragmentation ratio and poor MS/MS spectra. Several methods have been developed to circumvent this drawback. Derivatization of peptides with compounds containing positive charge has been studied; Chen et al. (RCMS, 2004, 18, 191) demonstrated the simplification of CID spectra of tryptic peptides modified by 4-sulfophenylisothiocyanate. The result is a predominance of y-type ions. In this work, we evaluate the potential of SPITC for the de novo sequencing of unknown non-tryptic peptides containing disulfide bridges, i.e. peptide toxins from animal venoms. Methods 2µL of peptide solution (100 µM) were diluted in 6µL NH4HCO3 50mM (pH 8.7). As peptide toxins often contain disulfide bridges, reduction (2µL DTT 50mM, 1h at 56°C) and alkylation (2µL IAA 500mM, 1h in darkness at RT) of peptides were performed before the derivatization reaction. Peptides were then adsorbed on a C18 ZipTip micro-column followed by 10 µL of 4-sulfophenylisothiocyanate (SPITC) 50mM. The column was then incubated for 6h at 56°C. Peptides were washed by TFA 0.2% and eluted in 10µL 50/50 ACN/FA 0.1%, before being spotted in 2,5-DHB. MS experiments were performed using a Bruker Ultraflex II MALDI-TOF/TOF. FlexControl 3.0, FlexAnalysis 3.0, BioTools 3.2 and SequenceEditor 3.2 softwares (Bruker Daltonics, Bremen) were used for data acquisition and interpretation. Preliminary data According to our first results, SPITC derivatization allows in positive mode to direct the fragmentation thanks to the acidic character of the sulfonate moiety present on the modified molecule. Indeed, a large series of y-type ions is found in the CID spectra allowing determining easily large sequence tags. Moreover, the number of C-terminus ions (b- and a-type ions) decreases, which improve the simplification of MS/MS spectra. Due to this fragmentation pattern, SPITC derivatization is clearly valuable for the sequencing of peptides that are not described in databases (de novo sequencing). For example, animal venoms are composed of several hundreds of peptides that are poorly studied, up to now. These peptides display a high importance for pharmaceutical applications and their sequencing is, as a consequence, of prime interest. Peptide toxins, which are not resulting from an enzymatic digestion, are however difficult to sequence by classical MS/MS methods. In this work, we demonstrate that the modification of peptide toxins with SPITC reagent is suitable for “real” de novo sequencing. The method was applied to isolated peptides as well as chromatographic fractions that contain up to 30 toxins. The perspectives of this work rest on the study of the SPITC modified peptides in negative mode. We expect to obtain a better sensitivity due to the presence of the negative sulfonic acid group at the N-terminus extremity, and also interesting MS/MS spectra including mainly a- or b-type ions. The final challenge will be the application of the protocol to high throughput sequencing of peptide toxins from a large variety of animal venoms. Novel aspect De novo sequencing of unusual non-tryptic peptides thanks to 4-sulfophenylisothiocyanate derivatization by post-source decay MALDI-MS [less ▲]

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See detailIsolation and cultivation of cellulolytic and xylanolytic bacteria and molds extracted from the gut of the termite Reticulitermes santonensis (3DV.1.14)
Tarayre, Cédric ULg; Bauwens, Julien ULg; Mattéotti, Christel et al

Poster (2013, June)

Biofuel production can be based on the use of agro-residues, consisting in a complex lignocellulosic structure which is not easily hydrolysable. The digestive tract of the termite Reticulitermes ... [more ▼]

Biofuel production can be based on the use of agro-residues, consisting in a complex lignocellulosic structure which is not easily hydrolysable. The digestive tract of the termite Reticulitermes santonensis contains a diversified microflora able to hydrolyze the wood components. Bacteria, molds and protists form efficient consortia, able to break the lignocellulosic complex by producing enzymes, such as xylanases and cellulases. Our purpose is the isolation of microbial strains from termite guts in order to evaluate their potential for hydrolysis of lignocellulosic materials. Termites were fed using different diets chosen to improve the xylanolytic and cellulolytic microflora: wood, microcristalline cellulose (added with lignin or not), α-cellulose (added with lignin or not) and birchwood xylan. Then, dissections were realized to isolate the potential xylanolytic and cellulolytic strains. This approach led us to isolate and to study several strains of bacteria (Bacillus sp. strain CTGx and Chryseobacterium sp. strain CTGx) and molds (Trichoderma virens strain CTGx and Sarocladium kiliense strain CTGx). These microorganisms were able to hydrolyze starch, xylan, cellulose, carboxymethylcellulose, esculin, β-glucan and Whatman® filter paper. They can produce glucose and xylose monomers and oligomers which can be further fermented to produce bioethanol. [less ▲]

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See detailResearch of New Enzyme Producing Strains in the Gut of the Termite Reticulitermes santonensis
Tarayre, Cédric ULg; Bauwens, Julien ULg; Mattéotti, Christel et al

Poster (2013, June)

Termites contain a complex microflora inside of their guts. Inferior termites contain bacteria, mycetes and protists that interact to degrade vegetable components. These strains act as consortia to break ... [more ▼]

Termites contain a complex microflora inside of their guts. Inferior termites contain bacteria, mycetes and protists that interact to degrade vegetable components. These strains act as consortia to break natural materials by secreting various enzymes. Our aim was the isolation and cultivation of microorganisms in order to produce new enzymes that can be further used in green chemistry. Termites were fed with different diets: pinewood, microcristalline cellulose (added with lignin or not), α-cellulose (added with lignin or not) and birchwood xylan. Then, dissections were realized to isolate interesting strains. All the microorganisms were subjected to enzyme assays. That technique allowed us to isolate and to cultivate various strains of bacteria, molds and protists. Three strains of bacteria, two strains of molds and one strain of protist were isolated and displayed different enzymatic activities. The bacteria Bacillus subtilis strain ABGx, Bacillus sp. strain CTGx and Chryseobacterium sp. strain CTGx displayed amylase, cellulase and xylanase activities. The molds Trichoderma virens strain CTGx and Sarocladium kiliense strain CTGx were also able to produce those enzymes. However, the protist Poterioochromonas sp. was found to produce only amylase. In conlusion, the termite gut is a complex culivation medium that provides a habitat for many microorganisms that show interesting enzymatic activities. [less ▲]

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See detailDevelopment of a quantitative approach to measure phospholipids in dried drops by Raman spectroscopy
Malherbe, Cédric ULg; Jadoul, Laure ULg; Gilbert, Bernard ULg et al

Poster (2013, May 24)

Phospholipids, PL, such as the phosphatidylcholine PC(18:0/18:1), play a role in the structure of living cells and are suspected to be part of the development of some diseases, for example cancers. Mass ... [more ▼]

Phospholipids, PL, such as the phosphatidylcholine PC(18:0/18:1), play a role in the structure of living cells and are suspected to be part of the development of some diseases, for example cancers. Mass spectrometry enables the structural analysis of PL in complex biological media but imaging mass spectrometry by MALDI-MS is rather limited for quantification purposes. Complementarily, Raman spectroscopy as a non invasive and non destructive method is a potential candidate to quantify and visualise the spatial distribution of the PL by molecular imaging. Unfortunately, the lack of specific chemical function in PL, compared to others biomolecules, limits the use of Raman spectroscopy in the identification process of those PL in complex biological samples. The results presented here belong to a first study of the application of the Raman analyses on dried residues of PL and mice brain tissue performed in the lab. [less ▲]

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See detailMechanism of resistance to anti-angiogenic therapy.
Cimino, Jonathan ULg; Sounni, Nor Eddine ULg; Debois, Delphine ULg et al

Poster (2013, May 17)

Detailed reference viewed: 8 (1 ULg)
See detailNew mechanism of resistance to anti-angiogenic therapy.
Cimino, Jonathan ULg; Sounni, Nor Eddine ULg; Debois, Delphine ULg et al

Conference (2013, May 13)

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See detailLes venins d'animaux, nouvelle panacée?
Echterbille, Julien ULg; Quinton, Loïc ULg; De Pauw, Edwin ULg

in Athena (2013)

Araignées, serpents, scorpions,… autant d’animaux ayant une place particulière dans l’imaginaire collectif. Fascinants, horripilants voire même terrifiants, les adjectifs ne manquent pas pour qualifier ... [more ▼]

Araignées, serpents, scorpions,… autant d’animaux ayant une place particulière dans l’imaginaire collectif. Fascinants, horripilants voire même terrifiants, les adjectifs ne manquent pas pour qualifier les réactions qu’ils suscitent auprès des populations. Que dire alors de la peur engendrée par leur venin. Une simple piqûre, morsure ou contact peut s’avérer extrêmement dangereux voire létal... [less ▲]

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See detailIntraocular lenses with functionalized surfaces by biomolecules in relation with lens epithelial cell adhesion
Huang, Yi-Shiang ULg; Alexandre, Michaël ULg; Bozukova, Dimitriya et al

Poster (2013, April 25)

A cataract is pathology opacity of the lens or capsule of the eye, causing impairment of vision or even blindness. Surgery, with lens extraction and intraocular lens implantation, is still the only ... [more ▼]

A cataract is pathology opacity of the lens or capsule of the eye, causing impairment of vision or even blindness. Surgery, with lens extraction and intraocular lens implantation, is still the only currently available treatment. The most common complication after implantation of intraocular lenses (IOLs) is the posterior capsular opacification (PCO) or secondary cataract. This is the result of lens epithelial cells (LECs) proliferation and their transition to mesenchymal cells. In 1997, a Sandwich theory was proposed to elucidate the developmental process of PCO. [1] According to this model, an IOL with higher affinity to LECs will induce a less PCO. In our research, the pHEMA (Poly(2-hydroxyethyl methacrylate)) based acrylic hydrophilic polymer is subjected to the surface modification by conjugating with the bioactive peptides. The RGD sequence, known for its excellent biocompatibility, is designed to stimulate the biointegration between the LECs and the polymer implant. [2]. From our research, The RGD peptide immobilized onto pHEMA surfaces significantly facilitates the adhesion of the porcine LEC. The peptide immobilized surface retains its biological function even after 10 times of autoclave. On the other hand, the immobilized peptide does not alter the hydrophobicity of the surface, the light transmission, as well as the cytotoxicity of the material. This functionalized biomaterial would possibly prevent the formation of PCO. [1] J Cataract Refract Surg. 1997 Dec;23(10):1539-42 [2] Trends Biotechnol. 2008 Jul;26(7):382-92 [less ▲]

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See detailAdvances in proteomics for the FP7 Venomics project
Degueldre, Michel ULg; Quinton, Loïc ULg; De Pauw, Edwin ULg

Scientific conference (2013, April)

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See detailDevelopment of an analytical method to detect metabolites of nitrofurans. Application to the study of furazolidone elimination in Vietnamese black tiger shrimp (Penaeus monodon)
Douny, Caroline ULg; Widart, Joëlle ULg; De Pauw, Edwin ULg et al

in Aquaculture (2013), 376-379

Nitrofurans, banned antibiotics in European Union (EU), have often been found in imported aquaculture products in the past and are still found nowadays according to the Rapid Alert System for Feed and ... [more ▼]

Nitrofurans, banned antibiotics in European Union (EU), have often been found in imported aquaculture products in the past and are still found nowadays according to the Rapid Alert System for Feed and Food (RASFF) of the European Commission. A quantitative method based on liquid chromatography coupled to isotopic dilution tandem mass spectrometry (LC–IDMS/MS) was developed for the determination of the residues of four nitrofuran antibiotic residues in shrimps. The experimental protocol consisted of an acid-catalysed release of protein-bound metabolites, followed by derivatisation with 2-nitrobenzaldehyde (NBA). Then, a double liquid–liquid extraction with ethyl acetate was performed before LC–IDMS/MS analysis by positive electrospray ionisation (ES+) with multiple reaction monitoring (MRM) of two transitions per compound. An “in-house” validation of the method for shrimp analysis was conducted according to the EU criteria for the analysis of veterinary drug residues in foods. The decision limits (CCalpha) were 0.08–0.36 µg kg -1 and the detection capabilities (CCbeta) were 0.12–0.61 µg kg-1, which are both below the minimum required performance limit (MRPL) set at 1 µg kg -1 by the EU. The developed method was applied to evaluate the elimination of furazolidone residues in shrimp muscles after a contamination experiment. After 28 days of decontamination, a concentration of 115 µg kg-1 of furazolidone metabolite 3-amino-2-oxazolidinone (AOZ) was still measured in shrimp muscle. [less ▲]

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See detailDiscrimination of Isobaric Leu/Ile Residues by MALDI In-source Decay Mass Spectrometry
Asakawa, Daiki; Smargiasso, Nicolas ULg; De Pauw, Edwin ULg

in Journal of the American Society for Mass Spectrometry (2013), 24(2), 297-300

MALDI in-source decay (ISD) has been used for the top-down sequencing of proteins. The use of 1,5-diaminonapthalene (1,5-DAN) gave strong intensity of w ions, which are informative fragments and can be ... [more ▼]

MALDI in-source decay (ISD) has been used for the top-down sequencing of proteins. The use of 1,5-diaminonapthalene (1,5-DAN) gave strong intensity of w ions, which are informative fragments and can be helpful for the distinction of the isobaric amino acids, Leu and Ile. Our data suggests that the w fragments are formed from z* radical fragment by unimolecular dissociation and high abundance of w ions in MALDI-ISD with 1,5-DAN can be understood as resulting from the low collision rate in the MALDI plume. The MALDI-ISD with 1,5-DAN could be a useful method for the top-down sequencing of proteins including discrimination of Leu and Ile near the C-terminal end. [less ▲]

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