title: Salt bridges regulate both dimer formation and monomeric flexibility in HdeB and may have a role in periplasmic chaperone function.
abstract: Escherichia coli and Gram-negative bacteria that live in the human gut must be able to tolerate rapid and large changes in environmental pH. Low pH irreversibly denatures and precipitates many bacterial proteins. While cytoplasmic proteins are well buffered against such swings, periplasmic proteins are not. Instead, it appears that some bacteria utilize chaperone proteins that stabilize periplasmic proteins, preventing their precipitation. Two highly expressed and related proteins, HdeA and HdeB, have been identified as acid-activated chaperones. The structure of HdeA is known and a mechanism for activation has been proposed. In this model, dimeric HdeA dissociates at low pH, and the exposed dimeric interface binds exposed hydrophobic surfaces of acid-denatured proteins, preventing their irreversible aggregation. We now report the structure and biophysical characterization of the HdeB protein. The monomer of HdeB shares a similar structure with HdeA, but its dimeric interface is different in composition and spatial location. We have used fluorescence to study the behavior of HdeB as pH is lowered, and like HdeA, it dissociates to monomers. We have identified one of the key intersubunit interactions that controls pH-induced monomerization. Our analysis identifies a structural interaction within the HdeB monomer that is disrupted as pH is lowered, leading to enhanced structural flexibility.
authors.affiliation: Biomedical Sciences Research Complex, University of St Andrews, St Andrews, Fife KY16 9ST, UK.
authors.forename: Wenjian
authors.initials: W
authors.lastname: Wang
authors.forename: Tim
authors.initials: T
authors.lastname: Rasmussen
authors.forename: Amanda J
authors.initials: AJ
authors.lastname: Harding
authors.forename: Nuala A
authors.initials: NA
authors.lastname: Booth
authors.forename: Ian R
authors.initials: IR
authors.lastname: Booth
authors.forename: James H
authors.initials: JH
authors.lastname: Naismith
mesh_terms: D002850:Chromatography, Gel; D002942:Circular Dichroism; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D011487:Protein Conformation; D055503:Protein Multimerization; D013057:Spectrum Analysis
chemical_list: D029968:Escherichia coli Proteins; C084502:hdeB protein, E coli