SEC-SAXS and HDX-MS: A powerful combination. The case of the calcium-binding domain of a bacterial toxin

2017 ◽  
Vol 65 (1) ◽  
pp. 62-68 ◽  
Author(s):  
Darragh P. O'Brien ◽  
Sébastien Brier ◽  
Daniel Ladant ◽  
Dominique Durand ◽  
Alexandre Chenal ◽  
...  
Keyword(s):  
Calcium Binding ◽  
Binding Domain ◽  
Bacterial Toxin ◽  
Hdx Ms ◽  
Calcium Binding Domain

scholarly journals FRCaMP, a Red Fluorescent Genetically Encoded Calcium Indicator Based on Calmodulin from Schizosaccharomyces Pombe Fungus

2020 ◽  
Vol 22 (1) ◽  
pp. 111
Author(s):  
Oksana M. Subach ◽  
Natalia V. Barykina ◽  
Elizaveta S. Chefanova ◽  
Anna V. Vlaskina ◽  
Vladimir P. Sotskov ◽  
...  
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Hela Cells ◽  
Calcium Binding ◽  
Dynamic Range ◽  
Spectral Characteristics ◽  
Binding Domain ◽  
Calcium Transients ◽  
Neuronal Cultures ◽  
Decay Kinetics ◽  
Calcium Binding Domain

Red fluorescent genetically encoded calcium indicators (GECIs) have expanded the available pallet of colors used for the visualization of neuronal calcium activity in vivo. However, their calcium-binding domain is restricted by calmodulin from metazoans. In this study, we developed red GECI, called FRCaMP, using calmodulin (CaM) from Schizosaccharomyces pombe fungus as a calcium binding domain. Compared to the R-GECO1 indicator in vitro, the purified protein FRCaMP had similar spectral characteristics, brightness, and pH stability but a 1.3-fold lower ΔF/F calcium response and 2.6-fold tighter calcium affinity with Kd of 441 nM and 2.4–6.6-fold lower photostability. In the cytosol of cultured HeLa cells, FRCaMP visualized calcium transients with a ΔF/F dynamic range of 5.6, which was similar to that of R-GECO1. FRCaMP robustly visualized the spontaneous activity of neuronal cultures and had a similar ΔF/F dynamic range of 1.7 but 2.1-fold faster decay kinetics vs. NCaMP7. On electrically stimulated cultured neurons, FRCaMP demonstrated 1.8-fold faster decay kinetics and 1.7-fold lower ΔF/F values per one action potential of 0.23 compared to the NCaMP7 indicator. The fungus-originating CaM of the FRCaMP indicator version with a deleted M13-like peptide did not interact with the cytosolic environment of the HeLa cells in contrast to the metazoa-originating CaM of the similarly truncated version of the GCaMP6s indicator with a deleted M13-like peptide. Finally, we generated a split version of the FRCaMP indicator, which allowed the simultaneous detection of calcium transients and the heterodimerization of bJun/bFos interacting proteins in the nuclei of HeLa cells with a ΔF/F dynamic range of 9.4 and a contrast of 2.3–3.5, respectively.

scholarly journals Mutagenesis of the fourth calcium-binding domain of yeast calmodulin

1993 ◽  
Vol 268 (18) ◽  
pp. 13267-13273
Author(s):  
I. Matsuura ◽  
E. Kimura ◽  
K. Tai ◽  
M. Yazawa
Keyword(s):  
Calcium Binding ◽  
Binding Domain ◽  
Calcium Binding Domain

A novel GSN variant outside the G2 calcium‐binding domain associated with Amyloidosis of the Finnish type

2021 ◽  
Author(s):  
Sean Mullany ◽  
Emmanuelle Souzeau ◽  
Sonja Klebe ◽  
Tiger Zhou ◽  
Lachlan S. W. Knight ◽  
...  
Keyword(s):  
Calcium Binding ◽  
Binding Domain ◽  
Finnish Type ◽  
Calcium Binding Domain

Calsyntenin-1, a Proteolytically Processed Postsynaptic Membrane Protein with a Cytoplasmic Calcium-Binding Domain

2001 ◽  
Vol 17 (1) ◽  
pp. 151-166 ◽  
Author(s):  
Lorenz Vogt ◽  
Sabine P. Schrimpf ◽  
Virginia Meskenaite ◽  
Renato Frischknecht ◽  
Jochen Kinter ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Calcium Binding ◽  
Postsynaptic Membrane ◽  
Binding Domain ◽  
Cytoplasmic Calcium ◽  
Calcium Binding Domain
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