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PMID: 20071602 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Combined bimolecular fluorescence complementation and Forster resonance energy transfer reveals ternary SNARE complex formation in living plant cells.

Plant physiology ·Vol. 152 ·No. 3 ·2010-03-00 ·Pages 1135-47

Kwaaitaal M, Keinath NF, Pajonk S, Biskup C, Panstruga R

Abstract

Various fluorophore-based microscopic methods, comprising Förster resonance energy transfer (FRET) and bimolecular fluorescence complementation (BiFC), are suitable to study pairwise interactions of proteins in living cells. The analysis of interactions between more than two protein partners using these methods, however, remains difficult. In this study, we report the successful application of combined BiFC-FRET-fluorescence lifetime imaging microscopy and BiFC-FRET-acceptor photobleaching measurements to visualize the formation of ternary soluble N-ethylmaleimide-sensitive factor attachment receptor complexes in leaf epidermal cells. This method expands the repertoire of techniques to study protein-protein interactions in living plant cells by a procedure capable of visualizing simultaneously interactions between three fluorophore-tagged polypeptide partners.

MeSH Terms
Fluorescence Resonance Energy Transfer/methods Luminescent Proteins/metabolism Microscopy, Fluorescence/methods Plant Epidermis/cytology,metabolism Plant Leaves/metabolism Plant Proteins/metabolism Protein Interaction Mapping SNARE Proteins/metabolism
Chemicals
Luminescent Proteins Plant Proteins SNARE Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Kwaaitaal Mark
Max-Planck Institute for Plant Breeding Research, D-50829 Cologne, Germany.
Keinath Nana F
Pajonk Simone
Biskup Christoph
Panstruga Ralph
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Article Info
Journal
Plant physiology
Abbr.
Plant Physiol
ISSN
1532-2548
DOI
10.1104/pp.109.151142
Published
2010-03-00
Epub
2010-00-13
Pages
1135-47
Language
English
Region
United States
NLM ID
0401224
PMCID
PMC2832253
Subset
IM
External Links
PubMed source DOI: 10.1104/pp.109.151142 PMC Full Text View journal details
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