Fluorescence lifetime imaging of interactions between Golgi tethering factors and Small GTPases in plants

  • Osterrieder A
  • Carvalho C
  • Latijnhouwers M
 et al. 
  • 52


    Mendeley users who have this article in their library.
  • 32


    Citations of this article.


Peripheral tethering factors bind to small GTPases in order to obtain their correct location within the Golgi apparatus. Using fluorescence resonance energy transfer (FRET) and fluorescence lifetime imaging microscopy (FLIM) we visualized interactions between Arabidopsis homologues of tethering factors and small GTPases at the Golgi stacks in planta. Co-expression of the coiled-coil proteins AtGRIP and golgin candidate 5 (GC5) [TATA element modulatory factor (TMF)] and the putative post-Golgi tethering factor AtVPS52 fused to green fluorescent protein (GFP) with mRFP (monomeric red fluorescent protein) fusions to the small GTPases AtRab-H1(b), AtRab-H1(c) and AtARL1 resulted in reduced GFP lifetimes compared to the control proteins. Interestingly, we observed differences in GFP quenching between the different protein combinations as well as selective quenching of GFP-AtVPS52-labelled structures. The data presented here indicate that the FRET-FLIM technique should prove invaluable in assessing protein interactions in living plant cells at the organelle level.

Author-supplied keywords

  • FRET
  • Flim
  • GFP
  • Golgi
  • Small GTPase
  • Tethering factor

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document


  • Anne OsterriederOxford Brookes University

  • Claudine M. Carvalho

  • Maita Latijnhouwers

  • Jorunn Nergaard Johansen

  • Christopher Stubbs

  • Stanley Botchway

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free