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Fluorescent Protein–photoprotein Fusions and Their Applications in Calcium Imaging
Author(s) -
Bakayan Adil,
Domingo Beatriz,
Vaquero Cecilia F.,
Peyriéras Nadine,
Llopis Juan
Publication year - 2017
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/php.12682
Subject(s) - aequorin , photoprotein , aequorea victoria , green fluorescent protein , bioluminescence , fluorescence , fusion protein , organelle , biology , microbiology and biotechnology , biophysics , chemistry , biochemistry , recombinant dna , optics , physics , gene , intracellular
Abstract Calcium‐activated photoproteins, such as aequorin, have been used as luminescent Ca 2+ indicators since 1967. After the cloning of aequorin in 1985, microinjection was substituted by its heterologous expression, which opened the way for a widespread use. Molecular fusion of green fluorescent protein (GFP) to aequorin recapitulated the nonradiative energy transfer process that occurs in the jellyfish Aequorea victoria , from which these two proteins were obtained, resulting in an increase of light emission and a shift to longer wavelength. The abundance and location of the chimera are seen by fluorescence, whereas its luminescence reports Ca 2+ levels. GFP ‐aequorin is broadly used in an increasing number of studies, from organelles and cells to intact organisms. By fusing other fluorescent proteins to aequorin, the available luminescence color palette has been expanded for multiplexing assays and for in vivo measurements. In this report, we will attempt to review the various photoproteins available, their reported fusions with fluorescent proteins and their biological applications to image Ca 2+ dynamics in organelles, cells, tissue explants and in live organisms.