Structural highlights
Function[GFP_AEQVI] Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca(2+)-activated photoprotein aequorin. Publication Abstract from PubMedReversibly photo-switchable proteins are essential for many super-resolution fluorescence microscopic and optoacoustic imaging methods. However, they have yet to be used as sensors that measure the distribution of specific analytes at the nanoscale or in the tissues of live animals. Here we constructed the prototype of a photo-switchable Ca(2+) sensor based on GCaMP5G that can be switched with 405/488-nm light and describe its molecular mechanisms at the structural level, including the importance of the interaction of the core barrel structure of the fluorescent protein with the Ca(2+) receptor moiety. We demonstrate super-resolution imaging of Ca(2+) concentration in cultured cells and optoacoustic Ca(2+) imaging in implanted tumor cells in mice under controlled Ca(2+) conditions. Finally, we show the generalizability of the concept by constructing examples of photo-switching maltose and dopamine sensors based on periplasmatic binding protein and G-protein-coupled receptor-based sensors. Genetically encoded photo-switchable molecular sensors for optoacoustic and super-resolution imaging.,Mishra K, Fuenzalida-Werner JP, Pennacchietti F, Janowski R, Chmyrov A, Huang Y, Zakian C, Klemm U, Testa I, Niessing D, Ntziachristos V, Stiel AC Nat Biotechnol. 2021 Nov 29. pii: 10.1038/s41587-021-01100-5. doi:, 10.1038/s41587-021-01100-5. PMID:34845372[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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