Expression and association of the Yersinia pestis translocon proteins, YopB and YopD, are facilitated by nanolipoprotein particles

Matthew A. Coleman, Jenny A. Cappuccio, Craig D. Blanchette, Tingjuan Gao, Erins Arroyo, Angela K. Hinz, Feliza A. Bourguet, Brent Segelke, Paul D. Hoeprich, Thomas Huser, Ted A. Laurence, Vladimir Motin, Brett A. Chromy

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

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 coexpressed (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.

Original languageEnglish (US)
Article numbere0150166
JournalPLoS One
Volume11
Issue number3
DOIs
StatePublished - Mar 1 2016

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Yersinia pestis
Membrane Proteins
Proteins
proteins
Pathogens
cells
Membranes
porins
type III secretion system
Porins
atomic force microscopy
protein transport
pathogens
outer membrane proteins
Atomic Force Microscopy
Fluorescence Spectrometry
protein aggregates
Protein Transport
Immunoprecipitation
Liposomes

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Coleman, M. A., Cappuccio, J. A., Blanchette, C. D., Gao, T., Arroyo, E., Hinz, A. K., ... Chromy, B. A. (2016). Expression and association of the Yersinia pestis translocon proteins, YopB and YopD, are facilitated by nanolipoprotein particles. PLoS One, 11(3), [e0150166]. https://doi.org/10.1371/journal.pone.0150166

Expression and association of the Yersinia pestis translocon proteins, YopB and YopD, are facilitated by nanolipoprotein particles. / Coleman, Matthew A.; Cappuccio, Jenny A.; Blanchette, Craig D.; Gao, Tingjuan; Arroyo, Erins; Hinz, Angela K.; Bourguet, Feliza A.; Segelke, Brent; Hoeprich, Paul D.; Huser, Thomas; Laurence, Ted A.; Motin, Vladimir; Chromy, Brett A.

In: PLoS One, Vol. 11, No. 3, e0150166, 01.03.2016.

Research output: Contribution to journalArticle

Coleman, MA, Cappuccio, JA, Blanchette, CD, Gao, T, Arroyo, E, Hinz, AK, Bourguet, FA, Segelke, B, Hoeprich, PD, Huser, T, Laurence, TA, Motin, V & Chromy, BA 2016, 'Expression and association of the Yersinia pestis translocon proteins, YopB and YopD, are facilitated by nanolipoprotein particles', PLoS One, vol. 11, no. 3, e0150166. https://doi.org/10.1371/journal.pone.0150166
Coleman, Matthew A. ; Cappuccio, Jenny A. ; Blanchette, Craig D. ; Gao, Tingjuan ; Arroyo, Erins ; Hinz, Angela K. ; Bourguet, Feliza A. ; Segelke, Brent ; Hoeprich, Paul D. ; Huser, Thomas ; Laurence, Ted A. ; Motin, Vladimir ; Chromy, Brett A. / Expression and association of the Yersinia pestis translocon proteins, YopB and YopD, are facilitated by nanolipoprotein particles. In: PLoS One. 2016 ; Vol. 11, No. 3.
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