Kerruth, S; Coates, C; Dürst, CD; Oertner, TG; Török, K
(2019)
The kinetic mechanisms of fast-decay red-fluorescent genetically encoded calcium indicators.
J Biol Chem, 294 (11).
pp. 3934-3946.
ISSN 1083-351X
https://doi.org/10.1074/jbc.RA118.004543
SGUL Authors: Torok, Katalin
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Abstract
Genetically encoded calcium indicators (GECIs) are useful reporters of cell-signaling, neuronal, and network activities. We have generated novel fast variants and investigated the kinetic mechanisms of two recently developed red-fluorescent GECIs (RGECIs), mApple-based jRGECO1a and mRuby-based jRCaMP1a. In the formation of fluorescent jRGECO1a and jRCaMP1a complexes, calcium binding is followed by rate-limiting isomerization. However, fluorescence decay of calcium-bound jRGECO1a follows a different pathway from its formation: dissociation of calcium occurs first, followed by the peptide, similarly to GCaMP-s. In contrast, fluorescence decay of calcium-bound jRCaMP1a occurs by the reversal of the on-pathway: peptide dissociation is followed by calcium. The mechanistic differences explain the generally slower off-kinetics of jRCaMP1a-type indicators compared with GCaMP-s and jRGECO1a-type GECI: the fluorescence decay rate of f-RCaMP1 was 21 s−1, compared with 109 s−1 for f-RGECO1 and f-RGECO2 (37 °C). Thus, the CaM–peptide interface is an important determinant of the kinetic responses of GECIs; however, the topology of the structural link to the fluorescent protein demonstrably affects the internal dynamics of the CaM–peptide complex. In the dendrites of hippocampal CA3 neurons, f-RGECO1 indicates calcium elevation in response to a 100 action potential train in a linear fashion, making the probe particularly useful for monitoring large-amplitude, fast signals, e.g. those in dendrites, muscle cells, and immune cells.
Item Type: | Article | ||||||||
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Additional Information: | © 2019 Kerruth et al. Author's Choice—Final version open access under the terms of the Creative Commons CC-BY license (http://creativecommons.org/licenses/by/4.0). | ||||||||
Keywords: | biosensor, calcium, calcium imaging, fluorescence, kinetics, biosensor, calcium, calcium imaging, fluorescence, kinetics, 06 Biological Sciences, 11 Medical And Health Sciences, 03 Chemical Sciences, Biochemistry & Molecular Biology | ||||||||
SGUL Research Institute / Research Centre: | Academic Structure > Molecular and Clinical Sciences Research Institute (MCS) | ||||||||
Journal or Publication Title: | J Biol Chem | ||||||||
ISSN: | 1083-351X | ||||||||
Language: | eng | ||||||||
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Publisher License: | Creative Commons: Attribution 4.0 | ||||||||
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PubMed ID: | 30651353 | ||||||||
Go to PubMed abstract | |||||||||
URI: | https://openaccess.sgul.ac.uk/id/eprint/110567 | ||||||||
Publisher's version: | https://doi.org/10.1074/jbc.RA118.004543 |
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