Functional determination of calcium-binding sites required for the activation of inositol 1,4,5-trisphosphate receptors

Authors

Vikas Arige, University of Rochester
Lara E. Terry, University of Rochester
Larry E. Wagner, University of Rochester
Sundeep Malik, University of Rochester
Mariah R. Baker, The University of Texas, Houston
Guizhen Fan, The University of Texas, Houston
Suresh K. Joseph, Thomas Jefferson UniversityFollow
Irina I Serysheva, The University of Texas, Houston
David I. Yule, University of Rochester

Document Type

Article

Publication Date

9-19-2022

Comments

This article is the author's' final published version in PNAS, Volume 119, Issue 39, September 2022.

This published version is available at https://doi.org/10.1073/pnas.2209267119. Copyright © 2022 the Authors.

Abstract

Inositol 1,4,5-trisphosphate receptors (IP3Rs) initiate a diverse array of physiological responses by carefully orchestrating intracellular calcium (Ca2+) signals in response to various external cues. Notably, IP3R channel activity is determined by several obligatory factors, including IP3, Ca2+, and ATP. The critical basic amino acid residues in the N-terminal IP3-binding core (IBC) region that facilitate IP3 binding are well characterized. In contrast, the residues conferring regulation by Ca2+ have yet to be ascertained. Using comparative structural analysis of Ca2+-binding sites identified in two main families of intracellular Ca2+-release channels, ryanodine receptors (RyRs) and IP3Rs, we identified putative acidic residues coordinating Ca2+ in the cytosolic calcium sensor region in IP3Rs. We determined the consequences of substituting putative Ca2+ binding, acidic residues in IP3R family members. We show that the agonist-induced Ca2+ release, single-channel open probability (P0), and Ca2+ sensitivities are markedly altered when the negative charge on the conserved acidic side chain residues is neutralized. Remarkably, neutralizing the negatively charged side chain on two of the residues individually in the putative Ca2+-binding pocket shifted the Ca2+ required to activate IP3R to higher concentrations, indicating that these residues likely are a component of the Ca2+ activation site in IP3R. Taken together, our findings indicate that Ca2+ binding to a well-conserved activation site is a common underlying mechanism resulting in increased channel activity shared by IP3Rs and RyRs.

Recommended Citation

Arige, Vikas; Terry, Lara E.; Wagner, Larry E.; Malik, Sundeep; Baker, Mariah R.; Fan, Guizhen; Joseph, Suresh K.; Serysheva, Irina I; and Yule, David I., "Functional determination of calcium-binding sites required for the activation of inositol 1,4,5-trisphosphate receptors" (2022). Department of Pathology, Anatomy, and Cell Biology Faculty Papers. Paper 368.
https://jdc.jefferson.edu/pacbfp/368

Creative Commons License

This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

PubMed ID

36122240

Language

English