A5073138461- Glycosylation and Glycoproteins Research
- Genomics and Phylogenetic Studies
- Bacteriophages and microbial interactions
- Monoclonal and Polyclonal Antibodies Research
- Bacterial Genetics and Biotechnology
- Mass Spectrometry Techniques and Applications
- Carbohydrate Chemistry and Synthesis
- Protein purification and stability
- Salmonella and Campylobacter epidemiology
- Plant Micronutrient Interactions and Effects
- Viral Infectious Diseases and Gene Expression in Insects
- Advanced Proteomics Techniques and Applications
- Bacillus and Francisella bacterial research
- Analytical Chemistry and Chromatography
- Microfluidic and Capillary Electrophoresis Applications
- Enzyme Structure and Function
- Phytochemical Studies and Bioactivities
- Potato Plant Research
- Phytase and its Applications
- Fluorine in Organic Chemistry
- Plant Pathogens and Fungal Diseases
- Galectins and Cancer Biology
- Chemical Reactions and Mechanisms
- Synthesis and Catalytic Reactions
- Ubiquitin and proteasome pathways
National Research Council Canada
2012-2024
Centre National de la Recherche Scientifique
2020
Trinity College Dublin
2020
National Academies of Sciences, Engineering, and Medicine
2003-2014
Stetson University
2012
Institute for Biological Sciences
2001-2011
Royal College of Physicians of Edinburgh
2008
Naval Medical Research Command
2003
Zero to Three
2002
University of Ottawa
2002
The cellular alterations associated with skeletal muscle differentiation share a high degree of similarity key phenotypic changes usually ascribed to apoptosis. For example, actin fiber disassembly/reorganization is conserved feature both apoptosis and differentiating myoblasts the contractile protein, myosin light chain kinase, required for apoptotic membrane blebbing. As such, these observations suggest that induction in myogenic lineage may use overlapping mechanisms. Here, we report...
ABSTRACT Campylobacter jejuni remains the leading cause of bacterial gastroenteritis in developed countries, and yet little is known concerning mechanisms by which this fastidious organism survives within its environment. We have demonstrated that C. 11168 can form biofilms on a variety surfaces. Proteomic analyses planktonic biofilm-grown cells differences protein expression profiles between two growth modes. Proteins involved motility complex, including flagellins (FlaA, FlaB), filament...
Summary Analysis of the complete flagellin glycosylation locus Campylobacter jejuni strain 81–176 revealed a less complex genomic organization than corresponding region in genome strain, C. NCTC 11168. Twenty‐four 45 genes found between Cj1293 and Cj1337 11168 are missing 81–176. Mutation six new genes, addition to three previously reported, resulted non‐motile phenotype, consistent with role synthesis pseudaminic acid (PseAc) or transfer PseAc flagellin. Cj1316c pseA had been shown result...
We examined two variants of the genome-sequenced strain, Campylobacter jejuni NCTC11168, which show marked differences in their virulence properties including colonization poultry, invasion Caco-2 cells, and motility. Transcript profiles obtained from whole genome DNA microarrays proteome analyses demonstrated that these are reflected late flagellar structural components factors those involved glycosylation cytolethal distending toxin production. identified putative sigma(28) sigma(54)...
Summary Flagellins from Campylobacter jejuni 81‐176 and coli VC167 are heavily glycosylated. The major modifications on both flagellins pseudaminic acid (Pse5Ac7Ac), a nine carbon sugar that is similar to sialic acid, an acetamidino‐substituted analogue of (PseAm). Previous data have indicated PseAm synthesized via Pse5Ac7Ac in C. 81‐176, but the two sugars using independent pathways VC167. Cj1293 gene encodes putative UDP‐GlcNAc C 6 ‐dehydratase/C 4 ‐reductase protein required for...
While the concept of Quality-by-Design is addressed at upstream and downstream process development stages, we questioned whether there are advantages to addressing issues biologics quality early in design molecule based on fundamental biophysical characterization, thereby reduce complexities product stages. Although limited number bispecific therapeutics clinic, these developments have been plagued with difficulty producing materials sufficient quantity for both preclinical clinical studies....
The presence of α2,6-sialic acids on the Fc N-glycan provides anti-inflammatory properties to IgGs through a mechanism that remains unclear. Fc-sialylated are rare in humans as well industrial host cell lines such Chinese hamster ovary (CHO) cells. Facilitated access well-characterized α2,6-sialylated would help elucidate this intriguing IgG's effector function. This study presents method for efficient glycan α2,6-sialylation wild-type and F243A IgG1 mutant by transient co-expression with...
Abstract Protein expression from stably transfected Chinese hamster ovary (CHO) clones is an established but time‐consuming method for manufacturing therapeutic recombinant proteins. The use of faster, alternative approaches, such as non‐clonal stable pools, has been restricted due to lower productivity and longstanding regulatory guidelines. Recently, the performance pools improved dramatically, making them a viable option quickly producing drug substance GLP‐toxicology early‐phase clinical...
Francisella tularensis subsp. (type A) strain SCHU S4 is a prototypic of the pathogen that highly virulent for humans and other mammals. Its intradermal (i.d.) 50% lethal dose (LD50) mice <10 CFU. We discovered spontaneous mutant, designated FSC043, with an i.d. LD50 >10(8) FSC043 effectively vaccinated against challenge type A strain, protective efficacy was at least as good F. LVS, empirically attenuated which has been used efficacious human vaccine. Comparative proteomics to identify two...
Summary N ‐linked glycosylation is recognized as an important post‐translational modification across all three domains of life. However, the understanding genetic pathways for assembly and attachment glycans in eukaryotic bacterial systems far outweighs knowledge comparable processes Archaea. The recent characterization a novel trisaccharide [β‐Man p NAcA6Thr‐(1‐4)‐β‐Glc NAc3NAcA‐(1‐3)‐β‐Glc NAc] to asparagine residues Methanococcus voltae flagellin S‐layer proteins affords new opportunities...
Summary Flagellin from Campylobacter coli VC167 is post‐translationally modified at ≥ 16 amino acid residues with pseudaminic and three related derivatives. The predominant modification was 5,7‐diacetamido‐3,5,7,9 ‐ tetradeoxy l glycero manno nonulosonic (pseudaminic acid, Pse5Ac7Ac), a that has been described previously on flagellin jejuni 81‐176. lacked two modi‐fications present in 81‐176 instead had unique modifications of masses 431 432 Da. Flagellins both C. were also an acetamidino...
The flagellum of Methanococcus voltae is composed four structural flagellin proteins FlaA, FlaB1, FlaB2, and FlaB3. These possess a total 15 potential N-linked sequons (NX(S/T)) show mass shift on an SDS-polyacrylamide gel indicating significant post-translational modification. We describe here the characterization glycan from M. using spectrometry to examine proteolytic digests in combination with NMR analysis purified sensitive, cryogenically cooled probe. Nano-liquid chromatography-tandem...
ABSTRACT In eukaryotes, N-linked protein glycosylation is a universal modification involving addition of preformed oligosaccharides to select Asn-Xaa-Ser/Thr motifs and influencing multiple biological events. We recently demonstrated that Campylobacter jejuni the first member Bacteria possess an glycan pathway. this study, high-resolution magic angle spinning nuclear magnetic resonance (HR-MAS NMR) was applied probe quantitate C. N -glycan biosynthesis in vivo. To confirm HR-MAS NMR...
ABSTRACT Previously, we identified five genes (Cj1321 to Cj1326, of which Cj1325 and Cj1326 are a single gene) in the O-linked flagellin glycosylation island that highly prevalent Campylobacter jejuni isolates from chickens. We report mutagenesis, functional, structural data confirm this locus, Cj1324 particular, has significant contributory role colonization chickens by C. . A motile ΔCj1324 mutant with intact flagella was considerably less hydrophobic able autoagglutinate form biofilms...
Cerebral ischemia rapidly initiates structural and functional changes in brain vessels, including blood-brain barrier disruption, inflammation, angiogenesis. Molecular events that accompany these were investigated microvessels extracted using laser-capture microdissection (LCM) from Sprague-Dawley rats subjected to a 20 min transient global cerebral followed by 1, 6, or 24 h reperfusion. Proteins approximately 300 LCM captured (20-100 microm) ICAT-labeled analyzed nanoLC-MS. In-house...
The coupling of microfabricated devices to nanoelectrospray mass spectrometers using both a triple quadrupole and time-of-flight spectrometer (QqTOF MS) is presented for the analysis trace-level membrane proteins. Short disposable emitters were directly coupled chip device via low dead volume connection. analytical performance this integrated in terms sensitivity reproducibility was evaluated standard peptide mixtures. A concentration detection limit ranging from 3.2 43.5 nM different...
A novel method for the production of coated capillaries capillary zone electrophoresis−electrospray mass spectrometry (CZE−ESMS), operating at 100−200 nL/min flow rates is described. Electrophoretic conditions conducive to simultaneous operation ESMS were investigated microsprayers different inner diameters. The ruggedness and reliability CZE columns with 20 μm i.d. over 6 h uninterrupted operation. Detection limits in order 0.1−5 fmol obtained peptide standards molecular masses ranging from...
The application of microfabricated devices coupled to a quadrupole time-of-flight mass spectrometer (Qq-TOF-MS) is presented for the analysis trace level digests gel-isolated proteins. In order enhance sample loading proteomics analyses, two different on-chip preconcentration techniques were evaluated. First, stacking procedure that used polarity switching remove buffer prior zone electrophoresis was easily integrated on devices. With present chip design, this technique provided up 70 nL...