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Studies of the conformational stability of invasion plasmid antigen B from Shigella

Authors


  • Thermostability is a major aspect towards formulating a successful protein subunit vaccine. This article presents an assessment of the thermostability of IpaB, IpaB's cognate chaperone, IpgC and the protein-chaperone complex IpaB/IpgC from S. flexeri, using different spectroscopic techniques. To understand role of the cognate chaperone IpgC, the protective antigen IpaB and the chaperone were also studied individually. The results revealed greater thermostability of the proteins around neutral pH conditions. Furthermore the data can be used in designing screening studies for protein stabilizers.

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.

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