Biophysical and Stabilization Studies of the Chlamydia trachomatis Mouse Pneumonitis Major Outer Membrane Protein
- 3 August 2009
- journal article
- research article
- Published by American Chemical Society (ACS) in Molecular Pharmaceutics
- Vol. 6 (5), 1553-1561
- https://doi.org/10.1021/mp900110q
Abstract
Native Chlamydia trachomatis mouse pneumonitis major outer membrane protein (nMOMP) induces effective protection against genital infection in a mouse challenge model. The conformation of nMOMP is crucial to confer this protective immunity. To achieve a better understanding of the conformational behavior and stability of nMOMP, a number of spectroscopic techniques are employed to characterize the secondary structure (circular dichroism), tertiary structure (intrinsic fluorescence) and aggregation properties (static light scattering and optical density) as a function of pH (3−8) and temperature (10−87.5 °C). The data are summarized in an empirical phase diagram (EPD) which demonstrates that the thermal stability of nMOMP is strongly pH-dependent. Three distinctive regions are seen in the EPD. Below the major thermal transition regions, nMOMP remains in its native conformation over the pH range of 3−8. Above the thermal transitions, nMOMP appears in two different structurally altered states; one at pH 3−5 and the other at pH 6−8. The EPD shows that the highest thermal transition point (∼65 °C) of nMOMP is near pH 6. Several potential excipients such as arginine, sodium citrate, Brij 35, sucrose and guanidine are also selected to evaluate their effects on the stability of nMOMP. These particular compounds increase the aggregation onset temperature of nMOMP by more than 10οC, without affecting its secondary and tertiary structure. These results should help formulate a vaccine using a recombinant MOMP.Keywords
This publication has 31 references indexed in Scilit:
- Physical Characterization of Clostridium Difficile Toxins and Toxoids: Effect of the Formaldehyde Crosslinking on Thermal StabilityJournal of Pharmaceutical Sciences, 2008
- Structural and Functional Analyses of the Major Outer Membrane Protein ofChlamydia trachomatisJournal of Bacteriology, 2007
- Characterization of the Disulfide Bonds and Free Cysteine Residues of the Chlamydia trachomatis Mouse Pneumonitis Major Outer Membrane ProteinBiochemistry, 2005
- Prevalence of Chlamydial and Gonococcal Infections Among Young Adults in the United StatesJAMA, 2004
- Sexual health--health of the nationSexually Transmitted Infections, 2003
- Chlamydial Genomics and Vaccine Antigen DiscoveryThe Journal of Infectious Diseases, 2000
- The Potential for Vaccine Development against Chlamydial Infection and DiseaseThe Journal of Infectious Diseases, 2000
- Partial protection against genital reinfection by immunization of guinea-pigs with isolated outer-membrane proteins of the chlamydial agent of guinea-pig inclusion conjunctivitisJournal of General Microbiology, 1993
- Partially structured self-associating states of acidic fibroblast growth factorBiochemistry, 1993
- Effect of temperature and low pH on structure and stability of matrix porin in micellar detergent solutionsBiochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1986