The IP3 receptor/Ca2+ channel and its cellular function

2007 ◽  
Vol 74 ◽  
pp. 9-22 ◽  
Author(s):  
Katsuhiko Mikoshiba
Keyword(s):  
Exocrine Secretion ◽  
Axis Formation ◽  
Ip3 Receptor ◽  
Release Channel ◽  
Saliva Secretion ◽  
Binding Core ◽  
Gastric Juice Secretion ◽  
Trisphosphate Receptor ◽  
Type 3

The IP3R [IP3 (inositol 1,4,5-trisphosphate) receptor] is responsible for Ca2+ release from the ER (endoplasmic reticulum). We have been working extensively on the P400 protein, which is deficient in Purkinje-neuron-degenerating mutant mice. We have discovered that P400 is an IP3R and we have determined the primary sequence. Purified IP3R, when incorporated into a lipid bilayer, works as a Ca2+ release channel and overexpression of IP3R shows enhanced IP3 binding and channel activity. Addition of an antibody blocks Ca2+ oscillations indicating that IP3R1 works as a Ca2+ oscillator. Studies on the role of IP3R during development show that IP3R is involved in fertilization and is essential for determination of dorso-ventral axis formation. We found that IP3R is involved in neuronal plasticity. A double homozygous mutant of IP3R2 (IP3R type 2) and IP3R3 (IP3R type 3) shows a deficit of saliva secretion and gastric juice secretion suggesting that IP3Rs are essential for exocrine secretion. IP3R has various unique properties: cryo-EM (electron microscopy) studies show that IP3R contains multiple cavities; IP3R allosterically and dynamically changes its form reversibly (square form–windmill form); IP3R is functional even though it is fragmented by proteases into several pieces; the ER forms a meshwork but also forms vesicular ER and moves along microtubules using a kinesin motor; X ray analysis of the crystal structure of the IP3 binding core consists of an N-terminal β-trefoil domain and a C-terminal α-helical domain. We have discovered ERp44 as a redox sensor in the ER which binds to the luminal part of IP3R1 and regulates its activity. We have also found the role of IP3 is not only to release Ca2+ but also to release IRBIT which binds to the IP3 binding core of IP3R.

scholarly journals Permeant calcium ion feed-through regulation of single inositol 1,4,5-trisphosphate receptor channel gating

2012 ◽  
Vol 140 (6) ◽  
pp. 697-716 ◽  
Author(s):  
Horia Vais ◽  
J. Kevin Foskett ◽  
Ghanim Ullah ◽  
John E. Pearson ◽  
Don-On Daniel Mak
Keyword(s):  
Posttranslational Modifications ◽  
Imaging Techniques ◽  
Channel Gating ◽  
Channel Activity ◽  
Release Channel ◽  
Solution Exchange ◽  
Inositol 1,4,5 Trisphosphate ◽  
Direct Binding ◽  
Trisphosphate Receptor ◽  
Type 3

The ubiquitous inositol 1,4,5-trisphosphate (InsP3) receptor (InsP3R) Ca2+ release channel plays a central role in the generation and modulation of intracellular Ca2+ signals, and is intricately regulated by multiple mechanisms including cytoplasmic ligand (InsP3, free Ca2+, free ATP4−) binding, posttranslational modifications, and interactions with cytoplasmic and endoplasmic reticulum (ER) luminal proteins. However, regulation of InsP3R channel activity by free Ca2+ in the ER lumen ([Ca2+]ER) remains poorly understood because of limitations of Ca2+ flux measurements and imaging techniques. Here, we used nuclear patch-clamp experiments in excised luminal-side-out configuration with perfusion solution exchange to study the effects of [Ca2+]ER on homotetrameric rat type 3 InsP3R channel activity. In optimal [Ca2+]i and subsaturating [InsP3], jumps of [Ca2+]ER from 70 nM to 300 µM reduced channel activity significantly. This inhibition was abrogated by saturating InsP3 but restored when [Ca2+]ER was raised to 1.1 mM. In suboptimal [Ca2+]i, jumps of [Ca2+]ER (70 nM to 300 µM) enhanced channel activity. Thus, [Ca2+]ER effects on channel activity exhibited a biphasic dependence on [Ca2+]i. In addition, the effect of high [Ca2+]ER was attenuated when a voltage was applied to oppose Ca2+ flux through the channel. These observations can be accounted for by Ca2+ flux driven through the open InsP3R channel by [Ca2+]ER, raising local [Ca2+]i around the channel to regulate its activity through its cytoplasmic regulatory Ca2+-binding sites. Importantly, [Ca2+]ER regulation of InsP3R channel activity depended on cytoplasmic Ca2+-buffering conditions: it was more pronounced when [Ca2+]i was weakly buffered but completely abolished in strong Ca2+-buffering conditions. With strong cytoplasmic buffering and Ca2+ flux sufficiently reduced by applied voltage, both activation and inhibition of InsP3R channel gating by physiological levels of [Ca2+]ER were completely abolished. Collectively, these results rule out Ca2+ regulation of channel activity by direct binding to the luminal aspect of the channel.

scholarly journals Single-Channel Properties in Endoplasmic Reticulum Membrane of Recombinant Type 3 Inositol Trisphosphate Receptor

2000 ◽  
Vol 115 (3) ◽  
pp. 241-256 ◽  
Author(s):  
Don-On Daniel Mak ◽  
Sean McBride ◽  
Viswanathan Raghuram ◽  
Yun Yue ◽  
Suresh K. Joseph ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Single Channel ◽  
Calcium Influx ◽  
Endoplasmic Reticulum Membrane ◽  
Release Channel ◽  
Cellular Expression ◽  
Recombinant Type ◽  
Specific Regulation ◽  
Trisphosphate Receptor ◽  
Type 3

The inositol 1,4,5-trisphosphate receptor (InsP3R) is an intracellular Ca2+-release channel localized in endoplasmic reticulum (ER) with a central role in complex Ca2+ signaling in most cell types. A family of InsP3Rs encoded by several genes has been identified with different primary sequences, subcellular locations, variable ratios of expression, and heteromultimer formation. This diversity suggests that cells require distinct InsP3Rs, but the functional correlates of this diversity are largely unknown. Lacking are single-channel recordings of the recombinant type 3 receptor (InsP3R-3), a widely expressed isoform also implicated in plasma membrane Ca2+ influx and apoptosis. Here, we describe functional expression and single-channel recording of recombinant rat InsP3R-3 in its native membrane environment. The approach we describe suggests a novel strategy for expression and recording of recombinant ER-localized ion channels in the ER membrane. Ion permeation and channel gating properties of the rat InsP3R-3 are strikingly similar to those of Xenopus type 1 InsP3R in the same membrane. Using two different two-electrode voltage clamp protocols to examine calcium store-operated calcium influx, no difference in the magnitude of calcium influx was observed in oocytes injected with rat InsP3R-3 cRNA compared with control oocytes. Our results suggest that if cellular expression of multiple InsP3R isoforms is a mechanism to modify the temporal and spatial features of [Ca2+]i signals, then it must be achieved by isoform-specific regulation or localization of various types of InsP3Rs that have relatively similar Ca2+ permeation properties.

scholarly journals Calcium Signaling in Cholangiocytes: Methods, Mechanisms, and Effects

2018 ◽  
Vol 19 (12) ◽  
pp. 3913 ◽  
Author(s):  
Michele Rodrigues ◽  
Dawidson Gomes ◽  
Michael Nathanson
Keyword(s):  
Calcium Signaling ◽  
Second Messenger ◽  
Experimental Models ◽  
Cell Type ◽  
Release Channel ◽  
Cellular Processes ◽  
Inositol 1,4,5 Trisphosphate ◽  
Regulation Of Secretion ◽  
Trisphosphate Receptor

Calcium (Ca2+) is a versatile second messenger that regulates a number of cellular processes in virtually every type of cell. The inositol 1,4,5-trisphosphate receptor (ITPR) is the only intracellular Ca2+ release channel in cholangiocytes, and is therefore responsible for Ca2+-mediated processes in these cells. This review will discuss the machinery responsible for Ca2+ signals in these cells, as well as experimental models used to investigate cholangiocyte Ca2+ signaling. We will also discuss the role of Ca2+ in the normal and abnormal regulation of secretion and apoptosis in cholangiocytes, two of the best characterized processes mediated by Ca2+ in this cell type.

IP3 receptor-dependent Ca2+ release modulates excitation-contraction coupling in rabbit ventricular myocytes

2008 ◽  
Vol 294 (2) ◽  
pp. H596-H604 ◽  
Author(s):  
Timothy L. Domeier ◽  
Aleksey V. Zima ◽  
Joshua T. Maxwell ◽  
Sabine Huke ◽  
Gregory A. Mignery ◽  
...  
Keyword(s):  
Ventricular Myocytes ◽  
Ventricular Myocardium ◽  
Ip3 Receptor ◽  
Inotropic Effects ◽  
Excitation Contraction Coupling ◽  
Contraction Coupling ◽  
Rabbit Ventricular Myocytes ◽  
Receptor Clusters ◽  
Type 3

Inositol 1,4,5-trisphosphate (IP3) receptor (IP3R)-dependent Ca2+ signaling exerts positive inotropic, but also arrhythmogenic, effects on excitation-contraction coupling (ECC) in the atrial myocardium. The role of IP3R-dependent sarcoplasmic reticulum (SR) Ca2+ release in ECC in the ventricular myocardium remains controversial. Here we investigated the role of this signaling pathway during ECC in isolated rabbit ventricular myocytes. Immunoblotting of proteins from ventricular myocytes showed expression of both type 2 and type 3 IP3R at levels ∼3.5-fold less than in atrial myocytes. In permeabilized myocytes, direct application of IP3 (10 μM) produced a transient 21% increase in the frequency of Ca2+ sparks ( P < 0.05). This increase was accompanied by a 13% decrease in spark amplitude ( P < 0.05) and a 7% decrease in SR Ca2+ load ( P < 0.05) and was inhibited by IP3R antagonists 2-aminoethoxydiphenylborate (2-APB; 20 μM) and heparin (0.5 mg/ml). In intact myocytes endothelin-1 (100 nM) was used to stimulate IP3 production and caused a 38% ( P < 0.05) increase in the amplitude of action potential-induced (0.5 Hz, field stimulation) Ca2+ transients. This effect was abolished by the IP3R antagonist 2-APB (2 μM) or by using adenoviral expression of an IP3 affinity trap that buffers cellular IP3. Together, these data suggest that in rabbit ventricular myocytes IP3R-dependent Ca2+ release has positive inotropic effects on ECC by facilitating Ca2+ release through ryanodine receptor clusters.

scholarly journals Thimerosal stimulates Ca2+ flux through inositol 1,4,5-trisphosphate receptor type 1, but not type 3, via modulation of an isoform-specific Ca2+-dependent intramolecular interaction

2004 ◽  
Vol 381 (1) ◽  
pp. 87-96 ◽  
Author(s):  
Geert BULTYNCK ◽  
Karolina SZLUFCIK ◽  
Nael Nadif KASRI ◽  
Zerihun ASSEFA ◽  
Geert CALLEWAERT ◽  
...  
Keyword(s):  
Amino Acids ◽  
Conformational Changes ◽  
Intramolecular Interaction ◽  
Binding Activity ◽  
Release Channel ◽  
B Lymphoma Cells ◽  
Inositol 1,4,5 Trisphosphate ◽  
Adenophostin A ◽  
Binding Core ◽  
Trisphosphate Receptor

Thiol-reactive agents such as thimerosal have been shown to modulate the Ca2+-flux properties of IP3 (inositol 1,4,5-trisphosphate) receptor (IP3R) via an as yet unidentified mechanism [Parys, Missiaen, De Smedt, Droogmans and Casteels (1993) Pflügers Arch. 424, 516–522; Kaplin, Ferris, Voglmaier and Snyder (1994) J. Biol. Chem. 269, 28972–28978; Missiaen, Taylor and Berridge (1992) J. Physiol. (Cambridge, U.K.) 455, 623–640; Missiaen, Parys, Sienaert, Maes, Kunzelmann, Takahashi, Tanzawa and De Smedt (1998) J. Biol. Chem. 273, 8983–8986]. In the present study, we show that thimerosal potentiated IICR (IP3-induced Ca2+ release) and IP3-binding activity of IP3R1, expressed in triple IP3R-knockout R23-11 cells derived from DT40 chicken B lymphoma cells, but not of IP3R3 or [Δ1–225]-IP3R1, which lacks the N-terminal suppressor domain. Using a 45Ca2+-flux technique in permeabilized A7r5 smooth-muscle cells, we have shown that Ca2+ shifted the stimulatory effect of thimerosal on IICR to lower concentrations of thimerosal and thereby increased the extent of Ca2+ release. This suggests that Ca2+ and thimerosal synergetically regulate IP3R1. Glutathione S-transferase pull-down experiments elucidated an interaction between amino acids 1–225 (suppressor domain) and amino acids 226–604 (IP3-binding core) of IP3R1, and this interaction was strengthened by both Ca2+ and thimerosal. In contrast, calmodulin and sCaBP-1 (short Ca2+-binding protein-1), both having binding sites in the 1–225 region, weakened the interaction. This interaction was not found for IP3R3, in agreement with the lack of functional stimulation of this isoform by thimerosal. The interaction between the IP3-binding and transmembrane domains (amino acids 1–604 and 2170–2749 respectively) was not affected by thimerosal and Ca2+, but it was significantly inhibited by IP3 and adenophostin A. Our results demonstrate that thimerosal and Ca2+ induce isoform-specific conformational changes in the N-terminal part of IP3R1, leading to the formation of a highly IP3-sensitive Ca2+-release channel.

scholarly journals The cardiac IP3 receptor: Uncovering the role of “the other” calcium-release channel

2008 ◽  
Vol 45 (2) ◽  
pp. 159-161 ◽  
Author(s):  
Thomas J. Hund ◽  
Andrew P. Ziman ◽  
W.J. Lederer ◽  
Peter J. Mohler
Keyword(s):  
Calcium Release ◽  
The Other ◽  
Ip3 Receptor ◽  
Calcium Release Channel ◽  
Release Channel

Centrosome Aurora A gradient ensures single polarity axis in C. elegans embryos

2020 ◽  
Vol 48 (3) ◽  
pp. 1243-1253 ◽  
Author(s):  
Sukriti Kapoor ◽  
Sachin Kotak
Keyword(s):  
Symmetry Breaking ◽  
Cell Fate ◽  
Cell Cortex ◽  
Axis Formation ◽  
Aurora A Kinase ◽  
Aurora A ◽  
C Elegans ◽  
Cell Embryo ◽  
Polarity Establishment

Cellular asymmetries are vital for generating cell fate diversity during development and in stem cells. In the newly fertilized Caenorhabditis elegans embryo, centrosomes are responsible for polarity establishment, i.e. anterior–posterior body axis formation. The signal for polarity originates from the centrosomes and is transmitted to the cell cortex, where it disassembles the actomyosin network. This event leads to symmetry breaking and the establishment of distinct domains of evolutionarily conserved PAR proteins. However, the identity of an essential component that localizes to the centrosomes and promotes symmetry breaking was unknown. Recent work has uncovered that the loss of Aurora A kinase (AIR-1 in C. elegans and hereafter referred to as Aurora A) in the one-cell embryo disrupts stereotypical actomyosin-based cortical flows that occur at the time of polarity establishment. This misregulation of actomyosin flow dynamics results in the occurrence of two polarity axes. Notably, the role of Aurora A in ensuring a single polarity axis is independent of its well-established function in centrosome maturation. The mechanism by which Aurora A directs symmetry breaking is likely through direct regulation of Rho-dependent contractility. In this mini-review, we will discuss the unconventional role of Aurora A kinase in polarity establishment in C. elegans embryos and propose a refined model of centrosome-dependent symmetry breaking.

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