Allosteric regulators selectively prevent Ca2+-feedback of CaV and NaV channels
Abstract
Calmodulin (CaM) serves as a pervasive regulatory subunit of CaV1, CaV2, and NaV1 channels, exploiting a functionally-conserved carboxy-tail element to afford dynamic Ca2+-feedback of cellular excitability in neurons and cardiomyocytes. Yet this modularity counters functional adaptability, as global changes in ambient CaM indiscriminately alter its targets. Here, we demonstrate that two structurally-unrelated proteins, SH3 and cysteine-rich domain (stac) and fibroblast growth factor homologous factors (fhf) selectively diminish Ca2+/CaM-regulation of CaV1 and NaV1 families respectively. The two proteins operate on allosteric sites within upstream portions of respective channel carboxy-tails, distinct from CaM-binding interface. Generalizing this mechanism, insertion of a short RxxK binding motif into CaV1.3 carboxy-tail, confers synthetic switching of CaM regulation by Mona SH3 domain. Overall, our findings identify a general class of auxiliary proteins that modify Ca2+/CaM signaling to individual targets allowing spatial and temporal orchestration of feedback, and outline strategies for engineering Ca2+/CaM signaling to individual targets.
Data availability
All data generated or analysed during this study are included in the manuscript and supporting files.
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Author details
Funding
National Institute of Mental Health
- David T Yue
- Manu Ben-Johny
National Heart, Lung, and Blood Institute
- Gordon Tomaselli
- Manu Ben-Johny
National Science Foundation
- Jacqueline Niu
National Institute of Neurological Disorders and Stroke
- Ivy E Dick
- David T Yue
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols of the Johns Hopkins University (GP15M172). The protocol was approved by the Committee on the Ethics of Animal Experiments of the Johns Hopkins University. All surgery was performed under sodium pentobarbital anesthesia, and every effort was made to minimize suffering.
Copyright
© 2018, Niu et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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