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ER-luminal [Ca²⁺] regulation of InsP₃ receptor gating mediated by an ER-luminal peripheral Ca²⁺-binding protein.

ABSTRACT: Modulating cytoplasmic Ca²⁺ concentration ([Ca²⁺]_i) by endoplasmic reticulum (ER)-localized inositol 1,4,5-trisphosphate receptor (InsP₃R) Ca²⁺-release channels is a universal signaling pathway that regulates numerous cell-physiological processes. Whereas much is known regarding regulation of InsP₃R activity by cytoplasmic ligands and processes, its regulation by ER-luminal Ca²⁺ concentration ([Ca²⁺]_ER) is poorly understood and controversial. We discovered that the InsP₃R is regulated by a peripheral membrane-associated ER-luminal protein that strongly inhibits the channel in the presence of high, physiological [Ca²⁺]_ER. The widely-expressed Ca²⁺-binding protein annexin A1 (ANXA1) is present in the nuclear envelope lumen and, through interaction with a luminal region of the channel, can modify high-[Ca²⁺]_ER inhibition of InsP₃R activity. Genetic knockdown of ANXA1 expression enhanced global and local elementary InsP₃-mediated Ca²⁺ signaling events. Thus, [Ca²⁺]_ER is a major regulator of InsP₃R channel activity and InsP₃R-mediated [Ca²⁺]_i signaling in cells by controlling an interaction of the channel with a peripheral membrane-associated Ca²⁺-binding protein, likely ANXA1.

SUBMITTER: Vais H

PROVIDER: S-EPMC7259957 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:The InsP3R Ca2+ release channel has a biphasic dependence on cytoplasmic free Ca2+ concentration ([Ca2+]i). InsP3 activates gating primarily by reducing the sensitivity of the channel to inhibition by high [Ca2+]i. To determine if relieving Ca2+ inhibition is sufficient for channel activation, we examined single-channel activities in low [Ca2+]i in the absence of InsP3, by patch clamping isolated Xenopus oocyte nuclei. For both endogenous Xenopus type 1 and recombinant rat type 3 InsP3R channels, spontaneous InsP3-independent channel activities with low open probability Po ( approximately 0.03) were observed in [Ca2+]i < 5 nM with the same frequency as in the presence of InsP3, whereas no activities were observed in 25 nM Ca2+. These results establish the half-maximal inhibitory [Ca2+]i of the channel to be 1.2-4.0 nM in the absence of InsP3, and demonstrate that the channel can be active when all of its ligand-binding sites (including InsP3) are unoccupied. In the simplest allosteric model that fits all observations in nuclear patch-clamp studies of [Ca2+]i and InsP3 regulation of steady-state channel gating behavior of types 1 and 3 InsP3R isoforms, including spontaneous InsP3-independent channel activities, the tetrameric channel can adopt six different conformations, the equilibria among which are controlled by two inhibitory and one activating Ca2+-binding and one InsP3-binding sites in a manner outlined in the Monod-Wyman-Changeux model. InsP3 binding activates gating by affecting the Ca2+ affinities of the high-affinity inhibitory sites in different conformations, transforming it into an activating site. Ca2+ inhibition of InsP3-liganded channels is mediated by an InsP3-independent low-affinity inhibitory site. The model also suggests that besides the ligand-regulated gating mechanism, the channel has a ligand-independent gating mechanism responsible for maximum channel Po being less than unity. The validity of this model was established by its successful quantitative prediction of channel behavior after it had been exposed to ultra-low bath [Ca2+].

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ER-luminal [Ca²⁺] regulation of InsP₃ receptor gating mediated by an ER-luminal peripheral Ca²⁺-binding protein.

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Dataset Information

ER-luminal [Ca2+] regulation of InsP3 receptor gating mediated by an ER-luminal peripheral Ca2+-binding protein.

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OmicsDI is part of the ELIXIR infrastructure

Tweets

ER-luminal [Ca²⁺] regulation of InsP₃ receptor gating mediated by an ER-luminal peripheral Ca²⁺-binding protein.