Cell-free production of a functional oligomeric form of a Chlamydia major outer-membrane protein (MOMP) for vaccine development
Abstract
Chlamydia is a prevalent sexually transmitted disease that infects more than 100 million people worldwide. Although most individuals infected with Chlamydia trachomatis are initially asymptomatic, symptoms can arise if left undiagnosed. Long-term infection can result in debilitating conditions such as pelvic inflammatory disease, infertility, and blindness. Chlamydia infection, therefore, constitutes a significant public health threat, underscoring the need for a Chlamydia-specific vaccine. Chlamydia strains express a major outer-membrane protein (MOMP) that has been shown to be an effective vaccine antigen. However, approaches to produce a functional recombinant MOMP protein for vaccine development are limited by poor solubility, low yield, and protein misfolding. For this study, we used an Escherichia coli-based cell-free system to express a MOMP protein from the mouse-specific species Chlamydia muridarum (MoPn-MOMP or mMOMP). The codon-optimized mMOMP gene was co-translated with Δ49apolipoprotein A1 (Δ49ApoA1), a truncated version of mouse ApoA1 in which the N-terminal 49 amino acids were removed. This co-translation process produced mMOMP supported within a telodendrimer nanolipoprotein particle (mMOMP–tNLP). The cell-free expressed mMOMP–tNLPs contain mMOMP multimers similar to the native MOMP protein. This cell-free process produced on average 1.5 mg of purified, water-soluble mMOMP–tNLP complex in a 1-ml cell-free reaction. The mMOMP–tNLP particle also accommodated themore »
- Authors:
-
- Lawrence Livermore National Laboratory (LLNL), Livermore, CA
- Synthetic Genomics Vaccines Inc. (SGVI), La Jolla, CA (United States)
- Lawrence Livermore National Laboratory (LLNL), Livermore, CA; Univ. of California, Merced, CA (United States). School of Natural Sciences
- Univ. of California, Davis, CA (United States). Dept. of Biochemistry and Molecular Medicine
- Univ. of California, Davis, CA (United States). Dept. of Molecular and Cellular Biology
- Lawrence Livermore National Laboratory (LLNL), Livermore, CA; Univ. of California, Davis, CA (United States). School of Medicine, Radiation Oncology
- Publication Date:
- Research Org.:
- Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
- Sponsoring Org.:
- USDOE; National Institutes of Health (NIH); Synthetic Genomics Vaccines Inc. (SGVI), La Jolla, CA (United States)
- OSTI Identifier:
- 1438621
- Report Number(s):
- LLNL-JRNL-747580
Journal ID: ISSN 0021-9258
- Grant/Contract Number:
- AC52-07NA27344; R01CA115483; R01CA140449; R21 RAI120925A
- Resource Type:
- Journal Article: Accepted Manuscript
- Journal Name:
- Journal of Biological Chemistry
- Additional Journal Information:
- Journal Volume: 292; Journal Issue: 36; Journal ID: ISSN 0021-9258
- Publisher:
- American Society for Biochemistry and Molecular Biology
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 60 APPLIED LIFE SCIENCES; 59 BASIC BIOLOGICAL SCIENCES; apolipoprotein; Chlamydia; membrane protein; nanotechnology; oligomer; cell-free expression; major outer membrane protein; nanolipoproteins; telodendrimer
Citation Formats
He, Wei, Felderman, Martina, Evans, Angela C., Geng, Jia, Homan, David, Bourguet, Feliza, Fischer, Nicholas O., Li, Yuanpei, Lam, Kit S., Noy, Aleksandr, Xing, Li, Cheng, R. Holland, Rasley, Amy, Blanchette, Craig D., Kamrud, Kurt, Wang, Nathaniel, Gouvis, Heather, Peterson, Todd C., Hubby, Bolyn, and Coleman, Matthew A. Cell-free production of a functional oligomeric form of a Chlamydia major outer-membrane protein (MOMP) for vaccine development. United States: N. p., 2017.
Web. doi:10.1074/jbc.M117.784561.
He, Wei, Felderman, Martina, Evans, Angela C., Geng, Jia, Homan, David, Bourguet, Feliza, Fischer, Nicholas O., Li, Yuanpei, Lam, Kit S., Noy, Aleksandr, Xing, Li, Cheng, R. Holland, Rasley, Amy, Blanchette, Craig D., Kamrud, Kurt, Wang, Nathaniel, Gouvis, Heather, Peterson, Todd C., Hubby, Bolyn, & Coleman, Matthew A. Cell-free production of a functional oligomeric form of a Chlamydia major outer-membrane protein (MOMP) for vaccine development. United States. https://doi.org/10.1074/jbc.M117.784561
He, Wei, Felderman, Martina, Evans, Angela C., Geng, Jia, Homan, David, Bourguet, Feliza, Fischer, Nicholas O., Li, Yuanpei, Lam, Kit S., Noy, Aleksandr, Xing, Li, Cheng, R. Holland, Rasley, Amy, Blanchette, Craig D., Kamrud, Kurt, Wang, Nathaniel, Gouvis, Heather, Peterson, Todd C., Hubby, Bolyn, and Coleman, Matthew A. 2017.
"Cell-free production of a functional oligomeric form of a Chlamydia major outer-membrane protein (MOMP) for vaccine development". United States. https://doi.org/10.1074/jbc.M117.784561. https://www.osti.gov/servlets/purl/1438621.
@article{osti_1438621,
title = {Cell-free production of a functional oligomeric form of a Chlamydia major outer-membrane protein (MOMP) for vaccine development},
author = {He, Wei and Felderman, Martina and Evans, Angela C. and Geng, Jia and Homan, David and Bourguet, Feliza and Fischer, Nicholas O. and Li, Yuanpei and Lam, Kit S. and Noy, Aleksandr and Xing, Li and Cheng, R. Holland and Rasley, Amy and Blanchette, Craig D. and Kamrud, Kurt and Wang, Nathaniel and Gouvis, Heather and Peterson, Todd C. and Hubby, Bolyn and Coleman, Matthew A.},
abstractNote = {Chlamydia is a prevalent sexually transmitted disease that infects more than 100 million people worldwide. Although most individuals infected with Chlamydia trachomatis are initially asymptomatic, symptoms can arise if left undiagnosed. Long-term infection can result in debilitating conditions such as pelvic inflammatory disease, infertility, and blindness. Chlamydia infection, therefore, constitutes a significant public health threat, underscoring the need for a Chlamydia-specific vaccine. Chlamydia strains express a major outer-membrane protein (MOMP) that has been shown to be an effective vaccine antigen. However, approaches to produce a functional recombinant MOMP protein for vaccine development are limited by poor solubility, low yield, and protein misfolding. For this study, we used an Escherichia coli-based cell-free system to express a MOMP protein from the mouse-specific species Chlamydia muridarum (MoPn-MOMP or mMOMP). The codon-optimized mMOMP gene was co-translated with Δ49apolipoprotein A1 (Δ49ApoA1), a truncated version of mouse ApoA1 in which the N-terminal 49 amino acids were removed. This co-translation process produced mMOMP supported within a telodendrimer nanolipoprotein particle (mMOMP–tNLP). The cell-free expressed mMOMP–tNLPs contain mMOMP multimers similar to the native MOMP protein. This cell-free process produced on average 1.5 mg of purified, water-soluble mMOMP–tNLP complex in a 1-ml cell-free reaction. The mMOMP–tNLP particle also accommodated the co-localization of CpG oligodeoxynucleotide 1826, a single-stranded synthetic DNA adjuvant, eliciting an enhanced humoral immune response in vaccinated mice. Using our mMOMP–tNLP formulation, we demonstrate a unique approach to solubilizing and administering membrane-bound proteins for future vaccine development. This method can be applied to other previously difficult-to-obtain antigens while maintaining full functionality and immunogenicity.},
doi = {10.1074/jbc.M117.784561},
url = {https://www.osti.gov/biblio/1438621},
journal = {Journal of Biological Chemistry},
issn = {0021-9258},
number = 36,
volume = 292,
place = {United States},
year = {Mon Jul 24 00:00:00 EDT 2017},
month = {Mon Jul 24 00:00:00 EDT 2017}
}
Web of Science
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