Bimolecular fluorescence complementation assay to explore protein-protein interactions of the Yersinia virulence factor YopM

Uğurlu Ö., EVRAN S.

Biochemical and Biophysical Research Communications, cilt.582, ss.43-48, 2021 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 582
  • Basım Tarihi: 2021
  • Doi Numarası: 10.1016/j.bbrc.2021.10.039
  • Dergi Adı: Biochemical and Biophysical Research Communications
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, Aquatic Science & Fisheries Abstracts (ASFA), BIOSIS, CAB Abstracts, Chemical Abstracts Core, EMBASE, Food Science & Technology Abstracts, MEDLINE, Veterinary Science Database
  • Sayfa Sayıları: ss.43-48
  • Anahtar Kelimeler: Effector protein, Fluorescence, Green fluorescent protein, Protein complementation, Protein interaction
  • Hatay Mustafa Kemal Üniversitesi Adresli: Evet

Özet

Yersinia outer protein M (YopM) is one of the effector proteins and essential for virulence. YopM is delivered by the Yersinia type III secretion system (T3SS) into the host cell, where it shows immunosuppressive effect through interaction with host proteins. Therefore, protein-protein interactions of YopM is significant to understand its molecular mechanism. In this study, we aimed to explore protein-protein interactions of YopM with the two components of T3SS, namely LcrV and LcrG. We used bimolecular fluorescence complementation (BiFC) assay and monitored the reassembly of green fluorescence protein in Escherichia coli. As an indicator of the protein-protein interaction, we monitored the in vivo reconstitution of fluorescence by measuring fluorescence intensity and imaging the cells under fluorescence microscope. We showed, for the first time, that YopM interacts with LcrG, but not with LcrV. Here, we propose BiFC assay as a simple method to screen novel interaction partners of YopM.