Spectroscopic analysis of chlamydial major outer membrane protein in support of structure elucidation

• DOI: 10.1002/pro.3501
• Publication Date: 2018-08-25T07:19:40Z
• Authors: Robert W. Hepler; Debbie D. Nahas; Bob Lucas; Robin Kaufhold; Jessica A Flynn; Jennifer D. Galli; Ryan Swoyer; James M. Wagner; Amy S. Espeseth; Joseph G. Joyce; James C. Cook; Eberhard Durr
• Keywords: 0301 basic medicine; Chlamydia muridarum; Full‐Length Papers; Circular Dichroism; Chlamydia trachomatis; Serogroup; 3. Good health; Cell Line; Protein Structure, Tertiary; Molecular Weight; 03 medical and health sciences; Cricetulus; Spectroscopy, Fourier Transform Infrared; Vaccines, Subunit; Animals; Humans; Protein Structure, Quaternary; Chromatography, High Pressure Liquid; Bacterial Outer Membrane Proteins

Abstract Chlamydial major outer membrane protein (MOMP) is the constituent of bacterial pathogen Chlamydia trachomatis. trachomatis Serovars D–K are leading cause genital tract infections which can lead to infertility or ectopic pregnancies. A vaccine against highly desirable but currently not available. MOMP accounts for ~ 60% chlamydial mass and considered be one candidates C. . We report on spectroscopic analysis native D, E, F, J as well muridarum by size exclusion chromatography multi angle light scattering (SEC MALS), circular dichroism (CD) attenuated total reflectance Fourier transform infrared spectroscopy (ATR‐FTIR). was purified from bacterium grown in either adherent HeLa cells different suspension cell lines. Our results confirm that forms homo‐trimers detergent micelles. The secondary structure composition conserved across serovars, with a 13% 18% (ATR‐FTIR) reduction β‐sheet conformation MOMP. When Serovar E isolated lines α‐helix content increased 7% (ATIR‐FTIR). Maintenance native‐like tertiary quaternary subunit vaccines important generation protective antibodies. This biophysical characterization presented here serves, absence functional assays, method monitoring structural integrity