PEB3 is a glycoprotein adhesin from Campylobacter jejuni whose structure suggested a role in transport. We have investigated potential ligands for PEB3 and characterized their binding properties using biophysical methods in solution and by X-ray crystallography. A thermal aggregation assay of PEB3 with a library of physiological compounds identified three possible ligands [3-phosphoglycerate (3-PG), phosphoenolpyruvate (PEP), and aconitate], which stabilized wild-type PEB3 but did not stabilize either a PEB3 form containing two mutations at the ligand-binding site, T138A/S139A, or a second PEB3 mutant, K135E, at a site approximately 14 A away. Fluorescence titration experiments and cocrystal structures with various ligands were used to characterize the binding of 3-PG, PEP, and phosphate to PEB3. Further, a C. jejuni growth experiment in minimal medium supplemented with 3-PG showed that this molecule enhances the growth of wild-type C. jejuni, but not of the PEB3 mutants. Crystallographic analysis of PEB3 complexes revealed that the Ser171-Gln180 region in the presence of 3-PG or other phosphates is helical and similar to those of other transport proteins, but it is nonhelical when citrate is bound. The K135E mutation resulted in expression of a more highly glycosylated form of PEB3 in vivo, and its crystal structure showed the conformation of the first two residues of the glycan. On the basis of our findings, we suggest that PEB3 is a transport protein that may function in utilization of 3-PG or other phosphate-containing molecules from the host.
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Front Psychol
June 2021
The Brown School, Washington University in St. Louis, St. Louis, MO, United States.
Policymakers are interested in programs that increase targeted pro-environmental behavior (PEB) and spill over to increase non-targeted PEBs. Theoretically, guilt should lead to negative spillover and identity to positive spillover, though this has rarely been tested empirically. Additionally, little is known about how reminders of past PEB behavior might also lead to downstream spillover effects.
View Article and Find Full Text PDFSci Rep
May 2018
Department of Pathogen Molecular Biology, London School of Hygiene and Tropical Medicine, Keppel Street, WC1E 7HT, London, UK.
Vibrio cholerae O1 El Tor is an aquatic Gram-negative bacterium responsible for the current seventh pandemic of the diarrheal disease, cholera. A previous whole-genome analysis on V. cholerae O1 El Tor strains from the 2010 epidemic in Pakistan showed that all strains contained the V.
View Article and Find Full Text PDFJ Biol Chem
October 2016
From the Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139
Site selectivity of protein N-linked glycosylation is dependent on many factors, including accessibility of the modification site, amino acid composition of the glycosylation consensus sequence, and cellular localization of target proteins. Previous studies have shown that the bacterial oligosaccharyltransferase, PglB, of Campylobacter jejuni favors acceptor proteins with consensus sequences ((D/E)XNX(S/T), where X ≠ proline) in flexible, solvent-exposed motifs; however, several native glycoproteins are known to harbor consensus sequences within structured regions of the acceptor protein, suggesting that unfolding or partial unfolding is required for efficient N-linked glycosylation in the native environment. To derive insight into these observations, we generated structural homology models of the N-linked glycoproteome of C.
View Article and Find Full Text PDFIn Silico Pharmacol
December 2016
Department of Biochemistry and Molecular Biology, University of Dhaka, Dhaka-, 1000, Bangladesh.
Purpose: Campylobacter jejuni is the one of the leading causes of bacterial diarrheal illness worldwide. This study aims to design specific epitopes for the utility of designing peptide vaccine(s) against C. jejuni by targeting invasive, virulent and membrane associated proteins like FlaA, Cia, CadF, PEB1, PEB3 and MOMP.
View Article and Find Full Text PDFBMC Microbiol
May 2014
School of Life Sciences, Kingston University, Faculty of Science, Engineering and Computing, Penrhyn Road, Kingston-upon Thames KT1 2EE, UK.
Background: Campylobacter jejuni (C. jejuni) is the leading causative agent of bacterial gastrointestinal infections. The rise of antibiotic resistant forms of this pathogen necessitates the development of novel intervention strategies.
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