title: Studies of the conformational stability of invasion plasmid antigen B from Shigella.
abstract: Shigella spp. are the causative agent of shigellosis, the second leading cause of diarrhea in children of ages 2-5. Despite many years of research, a protective vaccine has been elusive. We recently demonstrated that invasion plasmid antigens B and D (IpaB and IpaD) provide protection against S. flexneri and S. sonnei. These proteins, however, have very different properties which must be recognized and then managed during vaccine formulation. Herein, we employ spectroscopy to assess the stability of IpaB as well as IpgC (invasion protein gene), IpaB's cognate chaperone, and the IpaB/IpgC complex. The resulting data are mathematically summarized into a visual map illustrating the stability of the proteins and their complex as a function of pH and temperature. The IpaB/IpgC complex exhibits thermal stability at higher pH values but, though initially stable, quickly unfolds with increasing temperature when maintained at lower pH. In contrast, IpaB is a much more complex protein exhibiting increased stability at higher pH, but shows initial instability at lower pH values with pH 5 showing a distinct transition. IpgC precipitates at and below pH 5 and is stable above pH 7. Most strikingly, it is clear that complex formation results in stabilization of the two components. This work serves as a basis for the further development of IpaB as a vaccine candidate as well as extends our understanding of the structural stability of the Shigella type III secretion system.
authors.affiliation: Department of Microbiology and Molecular Genetics, Oklahoma State University, Stillwater, Oklahoma, USA.
authors.forename: Shyamal P
authors.initials: SP
authors.lastname: Choudhari
authors.forename: Ryan
authors.initials: R
authors.lastname: Kramer
authors.forename: Michael L
authors.initials: ML
authors.lastname: Barta
authors.forename: Jamie C
authors.initials: JC
authors.lastname: Greenwood
authors.forename: Brian V
authors.initials: BV
authors.lastname: Geisbrecht
authors.forename: Sangeeta B
authors.initials: SB
authors.lastname: Joshi
authors.forename: William D
authors.initials: WD
authors.lastname: Picking
authors.forename: C Russell
authors.initials: CR
authors.lastname: Middaugh
authors.forename: Wendy L
authors.initials: WL
authors.lastname: Picking
mesh_terms: D000942:Antigens, Bacterial / Q000737:chemistry* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D002648:Child; D002942:Circular Dichroism; D004405:Dysentery, Bacillary / Q000382:microbiology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D018832:Molecular Chaperones / Q000737:chemistry* / Q000378:metabolism; D055550:Protein Stability; D058767:Protein Unfolding; D012760:Shigella / Q000737:chemistry* / Q000378:metabolism
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D018832:Molecular Chaperones; C087447:ipaB protein, Shigella; C094580:IpgC protein, Shigella