Observation of the intracellular behavior of recombinant Yersinia pseudotuberculosis invasin protein

Hisashi Koga, Ichiro Izawa, Norie Araki, Hideyuki Saya, Tatsuyuki Mimori

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

In this study, we observed the intracellular behavior of recombinant invasin, a 103-kDa outer membrane protein of Yersinia pseudotuberculosis. To mimic the in vivo behavior of bacterial invasin, a polyvalent form of invasin was generated by incubation of biotinylated GST-fused invasin C-terminal portion protein (GST-INVS) with avidin. Several experiments confirmed that the recombinant invasin could consistently reproduce the invasin-mediated entry to mammalian epithelial cells. We analyzed the molecular kinetics of polyvalent INVS by western blotting, 125I-uptake, and immunofluorescent microscopy. The internalized polyvalent INVS was rapidly translocated to the RIPA-insoluble (polymerized-actin enriched) fraction and formed cytoplasmic vesicles, while monovalent invasin did not show such kinetics. From these observations, we concluded that our bacterial-free system is able to analyze the action of invasin for Yersinia pseudotuberculosis entry.

Original languageEnglish
Pages (from-to)297-302
Number of pages6
JournalMicrobiology and Immunology
Volume49
Issue number4
Publication statusPublished - 2005
Externally publishedYes

Fingerprint

Yersinia pseudotuberculosis
Cytoplasmic Vesicles
Avidin
Actins
Microscopy
Membrane Proteins
Proteins
Western Blotting
Epithelial Cells
Behavior Observation Techniques
Yersinia invasin

Keywords

  • Bacterial entry
  • Invasin
  • Yersinia pseudotuberculosis

ASJC Scopus subject areas

  • Immunology and Microbiology(all)
  • Microbiology (medical)
  • Microbiology

Cite this

Observation of the intracellular behavior of recombinant Yersinia pseudotuberculosis invasin protein. / Koga, Hisashi; Izawa, Ichiro; Araki, Norie; Saya, Hideyuki; Mimori, Tatsuyuki.

In: Microbiology and Immunology, Vol. 49, No. 4, 2005, p. 297-302.

Research output: Contribution to journalArticle

Koga, Hisashi ; Izawa, Ichiro ; Araki, Norie ; Saya, Hideyuki ; Mimori, Tatsuyuki. / Observation of the intracellular behavior of recombinant Yersinia pseudotuberculosis invasin protein. In: Microbiology and Immunology. 2005 ; Vol. 49, No. 4. pp. 297-302.
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