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PMID: 18155474 Published · ppublish English Journal Article

Bimolecular fluorescence complementation: visualization of molecular interactions in living cells.

Methods in cell biology ·Vol. 85 ·2008-00-00 ·Pages 431-70

Kerppola TK

Abstract

A variety of experimental methods have been developed for the analysis of protein interactions. The majority of these methods either require disruption of the cells to detect molecular interactions or rely on indirect detection of the protein interaction. The bimolecular fluorescence complementation (BiFC) assay provides a direct approach for the visualization of molecular interactions in living cells and organisms. The BiFC approach is based on the facilitated association between two fragments of a fluorescent protein when the fragments are brought together by an interaction between proteins fused to the fragments. The BiFC approach has been used for visualization of interactions among a variety of structurally diverse interaction partners in many different cell types. It enables detection of transient complexes as well as complexes formed by a subpopulation of the interaction partners. It is essential to include negative controls in each experiment in which the interface between the interaction partners has been mutated or deleted. The BiFC assay has been adapted for simultaneous visualization of multiple protein complexes in the same cell and the competition for shared interaction partners. A ubiquitin-mediated fluorescence complementation assay has also been developed for visualization of the covalent modification of proteins by ubiquitin family peptides. These fluorescence complementation assays have a great potential to illuminate a variety of biological interactions in the future.

MeSH Terms
Animals Fluorescence Luminescent Proteins Microscopy, Fluorescence/methods Proteins/metabolism Recombinant Fusion Proteins/metabolism Ubiquitins/metabolism
Chemicals
Luminescent Proteins Proteins Recombinant Fusion Proteins Ubiquitins
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Kerppola Tom K
Department of Biological Chemistry, Howard Hughes Medical Institute, University of Michigan Medical School, Ann Arbor, Michigan 48109, USA.
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Article Info
Journal
Methods in cell biology
Abbr.
Methods Cell Biol
ISSN
0091-679X
DOI
10.1016/S0091-679X(08)85019-4
Published
2008-00-00
Pages
431-70
Language
English
Region
United States
NLM ID
0373334
PMCID
PMC2829325
Subset
IM
Grants
Howard Hughes Medical Institute · United States
NIGMS NIH HHS · R01 GM086213 · United States
NIGMS NIH HHS · R01 GM086213-01 · United States
External Links
PubMed source DOI: 10.1016/S0091-679X(08)85019-4 PMC Full Text View journal details
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