Expression and Association of the Yersinia pestis Translocon Proteins, YopB and YopD, Are Facilitated by Nanolipoprotein Particles
| Autor: | Matthew A. Coleman, Tingjuan Gao, Erins Arroyo, Ted A. Laurence, Feliza Bourguet, Vladimir L. Motin, Paul D. Hoeprich, Brett A. Chromy, Angela K. Hinz, Jenny A. Cappuccio, Thomas R Huser, Craig D. Blanchette, Brent W. Segelke |
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| Přispěvatelé: | Skurnik, Mikael |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Yersinia pestis Cell Membranes lcsh:Medicine Pathology and Laboratory Medicine Microscopy Atomic Force Biochemistry Type three secretion system Medicine and Health Sciences 2.1 Biological and endogenous factors 2.2 Factors relating to the physical environment Aetiology lcsh:Science Gel Electrophoresis Microscopy Multidisciplinary biology Effector Vesicle Atomic Force Translocon Lipids Yersinia Cell biology Atomic Force Microscopy Bacterial Pathogens Infectious Diseases Medical Microbiology Cellular Structures and Organelles Pathogens Bacterial outer membrane Infection Research Article Bacterial Outer Membrane Proteins Yersinia Pestis Immunoprecipitation General Science & Technology Lipoproteins 030106 microbiology Research and Analysis Methods Microbiology Biophysical Phenomena Vaccine Related 03 medical and health sciences Electrophoretic Techniques Virology Biodefense Vesicles Microbial Pathogens Bacteria Scanning Probe Microscopy Prevention lcsh:R Host Cells Organisms Biology and Life Sciences Membrane Proteins Cell Biology biology.organism_classification Vector-Borne Diseases 030104 developmental biology Emerging Infectious Diseases Membrane protein Gene Expression Regulation Multiprotein Complexes Liposomes Nanoparticles lcsh:Q Viral Transmission and Infection |
| Zdroj: | PloS one, vol 11, iss 3 PLoS ONE PLoS ONE, Vol 11, Iss 3, p e0150166 (2016) |
| Popis: | Yersinia pestis enters host cells and evades host defenses, in part, through interactions between Yersinia pestis proteins and host membranes. One such interaction is through the type III secretion system, which uses a highly conserved and ordered complex for Yersinia pestis outer membrane effector protein translocation called the injectisome. The portion of the injectisome that interacts directly with host cell membranes is referred to as the translocon. The translocon is believed to form a pore allowing effector molecules to enter host cells. To facilitate mechanistic studies of the translocon, we have developed a cell-free approach for expressing translocon pore proteins as a complex supported in a bilayer membrane mimetic nano-scaffold known as a nanolipoprotein particle (NLP) Initial results show cell-free expression of Yersinia pestis outer membrane proteins YopB and YopD was enhanced in the presence of liposomes. However, these complexes tended to aggregate and precipitate. With the addition of co-expressed (NLP) forming components, the YopB and/or YopD complex was rendered soluble, increasing the yield of protein for biophysical studies. Biophysical methods such as Atomic Force Microscopy and Fluorescence Correlation Spectroscopy were used to confirm that the soluble YopB/D complex was associated with NLPs. An interaction between the YopB/D complex and NLP was validated by immunoprecipitation. The YopB/D translocon complex embedded in a NLP provides a platform for protein interaction studies between pathogen and host proteins. These studies will help elucidate the poorly understood mechanism which enables this pathogen to inject effector proteins into host cells, thus evading host defenses. |
| Databáze: | OpenAIRE |
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