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TitleDimerization-dependent green and yellow fluorescent proteins.
Publication TypeJournal Article
Year of Publication2012
AuthorsAlford, Spencer C., Yidan Ding, Thomas Simmen, and Robert E. Campbell
JournalACS Synth Biol
Volume1
Issue12
Pagination569-75
Date Published2012 Dec 21
ISSN2161-5063
KeywordsBacterial Proteins, Biosensing Techniques, Endoplasmic Reticulum, Fluorescent Antibody Technique, Green Fluorescent Proteins, Humans, Luminescent Proteins, Mitochondria, Mitochondrial Membranes, Protein Multimerization
Abstract

Dimerization-dependent fluorescent proteins (ddFP) are a recently introduced class of genetically encoded reporters that can be used for the detection of protein interactions in live cells. The progenitor of this class of tools was a red fluorescent ddFP (ddRFP) derived from a homodimeric variant of Discosoma red fluorescent protein. Here, we describe the engineering and application of an expanded palette of ddFPs, which includes green (ddGFP) and yellow (ddYFP) variants. These optimized variants offer several advantages relative to ddRFP including increased in vitro contrast and brightness for ddGFP and increased brightness and a lowered pK a for ddYFP. We demonstrate that both variants are useful as biosensors for protease activity in live cells. Using the ddGFP tool, we generated a highly effective indicator of endomembrane proximity that can be used to image the mitochondria-associated membrane (MAM) interface of endoplasmic reticulum (ER) and mitochondria.

DOI10.1021/sb300050j
Alternate JournalACS Synth Biol
PubMed ID23656278
PubMed Central IDPMC3653836
Grant List99085 / / Canadian Institutes of Health Research / Canada
/ / Canadian Institutes of Health Research / Canada