Specificity of the Campylobacter jejuni adhesin PEB3 for phosphates and structural differences among its ligand complexes

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DOIResolve DOI: http://doi.org/10.1021/bi802195d
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TypeArticle
Journal titleBiochemistry
Volume48
Issue14
Pages30573067; # of pages: 11
AbstractPEB3 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 ∼14 Å 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.
Publication date
LanguageEnglish
AffiliationNRC Institute for Biological Sciences; National Research Council Canada; NRC Biotechnology Research Institute
Peer reviewedYes
NPARC number12919054
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Record identifierb7c25009-f5f5-41c0-bfde-5c1819d50345
Record created2009-11-10
Record modified2016-05-09
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