Functional characterization of the type 1 inositol 1,4,5-trisphosphate receptor coupling domain SII(±) splice variants and the opisthotonos mutant form

Huiping Tu, Tomoya Miyakawa, Zhengnan Wang, Lyuba Glouchankova, Masamitsu Lino, Ilya Bezprozvanny

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

The type 1 inositol (1,4,5)-trisphosphate receptor (InsP3R1) plays a critical role in Ca2+ signaling in cells. Neuronal and nonneuronal isoforms of the InsP3R1 differ by alternative splicing in the coupling domain of the InsP3R1 (SII site) (Danoff et al., 1991). Deletion of 107 amino acids from the coupling domain of the InsP3R1 results in epileptic-like behaviors in opisthotonos (opt) spontaneous mouse mutant (Street et al., 1997). Using Spodoptera frugiperda cells expression system, we compared single-channel behavior of recombinant InsP3R1-SII(+), InsP3R1-SII(-), and InsP3R1-opt channels in planar lipid bilayers. The main results of our study are: 1) the InsP3R1-SII(-) has a higher conductance (94 pS) and the InsP3R1-opt has a lower conductance (64 pS) than the InsP3R1-SII(+) (81 pS); 2) the bell-shaped Ca2+-dependence peaks at 200-300 nM Ca2+ for all three InsP3R1 isoforms; 3) the bell-shaped Ca2+-dependence is wider for the InsP3R1-SII(+) and narrower for the InsP3R1-SII(-) and InsP3R1-opt; 4) the apparent affinity for ATP is sixfold lower for the InsP3R1-SII(-) (1.4 mM) and 20-fold lower for the InsP3R1-opt (5.3 mM) than for the InsP3R1-SII(+) (0.24 mM); 5) the InsP3R1-SII(-) is approximately twofold more active than the InsP3R1-SII(+) in the absence of ATP. Obtained results provide novel information about the molecular determinants of the InsP3R1 function.

Original languageEnglish (US)
Pages (from-to)1995-2004
Number of pages10
JournalBiophysical Journal
Volume82
Issue number4
StatePublished - 2002

Fingerprint

Inositol 1,4,5-Trisphosphate Receptors
Protein Isoforms
Adenosine Triphosphate
Spodoptera
Alternative Splicing
Lipid Bilayers
Amino Acids

ASJC Scopus subject areas

  • Biophysics

Cite this

Functional characterization of the type 1 inositol 1,4,5-trisphosphate receptor coupling domain SII(±) splice variants and the opisthotonos mutant form. / Tu, Huiping; Miyakawa, Tomoya; Wang, Zhengnan; Glouchankova, Lyuba; Lino, Masamitsu; Bezprozvanny, Ilya.

In: Biophysical Journal, Vol. 82, No. 4, 2002, p. 1995-2004.

Research output: Contribution to journalArticle

Tu, Huiping ; Miyakawa, Tomoya ; Wang, Zhengnan ; Glouchankova, Lyuba ; Lino, Masamitsu ; Bezprozvanny, Ilya. / Functional characterization of the type 1 inositol 1,4,5-trisphosphate receptor coupling domain SII(±) splice variants and the opisthotonos mutant form. In: Biophysical Journal. 2002 ; Vol. 82, No. 4. pp. 1995-2004.
@article{901a25e2f4dd4e7f9f0c84fb94875e2b,
title = "Functional characterization of the type 1 inositol 1,4,5-trisphosphate receptor coupling domain SII(±) splice variants and the opisthotonos mutant form",
abstract = "The type 1 inositol (1,4,5)-trisphosphate receptor (InsP3R1) plays a critical role in Ca2+ signaling in cells. Neuronal and nonneuronal isoforms of the InsP3R1 differ by alternative splicing in the coupling domain of the InsP3R1 (SII site) (Danoff et al., 1991). Deletion of 107 amino acids from the coupling domain of the InsP3R1 results in epileptic-like behaviors in opisthotonos (opt) spontaneous mouse mutant (Street et al., 1997). Using Spodoptera frugiperda cells expression system, we compared single-channel behavior of recombinant InsP3R1-SII(+), InsP3R1-SII(-), and InsP3R1-opt channels in planar lipid bilayers. The main results of our study are: 1) the InsP3R1-SII(-) has a higher conductance (94 pS) and the InsP3R1-opt has a lower conductance (64 pS) than the InsP3R1-SII(+) (81 pS); 2) the bell-shaped Ca2+-dependence peaks at 200-300 nM Ca2+ for all three InsP3R1 isoforms; 3) the bell-shaped Ca2+-dependence is wider for the InsP3R1-SII(+) and narrower for the InsP3R1-SII(-) and InsP3R1-opt; 4) the apparent affinity for ATP is sixfold lower for the InsP3R1-SII(-) (1.4 mM) and 20-fold lower for the InsP3R1-opt (5.3 mM) than for the InsP3R1-SII(+) (0.24 mM); 5) the InsP3R1-SII(-) is approximately twofold more active than the InsP3R1-SII(+) in the absence of ATP. Obtained results provide novel information about the molecular determinants of the InsP3R1 function.",
author = "Huiping Tu and Tomoya Miyakawa and Zhengnan Wang and Lyuba Glouchankova and Masamitsu Lino and Ilya Bezprozvanny",
year = "2002",
language = "English (US)",
volume = "82",
pages = "1995--2004",
journal = "Biophysical Journal",
issn = "0006-3495",
publisher = "Biophysical Society",
number = "4",

}

TY - JOUR

T1 - Functional characterization of the type 1 inositol 1,4,5-trisphosphate receptor coupling domain SII(±) splice variants and the opisthotonos mutant form

AU - Tu, Huiping

AU - Miyakawa, Tomoya

AU - Wang, Zhengnan

AU - Glouchankova, Lyuba

AU - Lino, Masamitsu

AU - Bezprozvanny, Ilya

PY - 2002

Y1 - 2002

N2 - The type 1 inositol (1,4,5)-trisphosphate receptor (InsP3R1) plays a critical role in Ca2+ signaling in cells. Neuronal and nonneuronal isoforms of the InsP3R1 differ by alternative splicing in the coupling domain of the InsP3R1 (SII site) (Danoff et al., 1991). Deletion of 107 amino acids from the coupling domain of the InsP3R1 results in epileptic-like behaviors in opisthotonos (opt) spontaneous mouse mutant (Street et al., 1997). Using Spodoptera frugiperda cells expression system, we compared single-channel behavior of recombinant InsP3R1-SII(+), InsP3R1-SII(-), and InsP3R1-opt channels in planar lipid bilayers. The main results of our study are: 1) the InsP3R1-SII(-) has a higher conductance (94 pS) and the InsP3R1-opt has a lower conductance (64 pS) than the InsP3R1-SII(+) (81 pS); 2) the bell-shaped Ca2+-dependence peaks at 200-300 nM Ca2+ for all three InsP3R1 isoforms; 3) the bell-shaped Ca2+-dependence is wider for the InsP3R1-SII(+) and narrower for the InsP3R1-SII(-) and InsP3R1-opt; 4) the apparent affinity for ATP is sixfold lower for the InsP3R1-SII(-) (1.4 mM) and 20-fold lower for the InsP3R1-opt (5.3 mM) than for the InsP3R1-SII(+) (0.24 mM); 5) the InsP3R1-SII(-) is approximately twofold more active than the InsP3R1-SII(+) in the absence of ATP. Obtained results provide novel information about the molecular determinants of the InsP3R1 function.

AB - The type 1 inositol (1,4,5)-trisphosphate receptor (InsP3R1) plays a critical role in Ca2+ signaling in cells. Neuronal and nonneuronal isoforms of the InsP3R1 differ by alternative splicing in the coupling domain of the InsP3R1 (SII site) (Danoff et al., 1991). Deletion of 107 amino acids from the coupling domain of the InsP3R1 results in epileptic-like behaviors in opisthotonos (opt) spontaneous mouse mutant (Street et al., 1997). Using Spodoptera frugiperda cells expression system, we compared single-channel behavior of recombinant InsP3R1-SII(+), InsP3R1-SII(-), and InsP3R1-opt channels in planar lipid bilayers. The main results of our study are: 1) the InsP3R1-SII(-) has a higher conductance (94 pS) and the InsP3R1-opt has a lower conductance (64 pS) than the InsP3R1-SII(+) (81 pS); 2) the bell-shaped Ca2+-dependence peaks at 200-300 nM Ca2+ for all three InsP3R1 isoforms; 3) the bell-shaped Ca2+-dependence is wider for the InsP3R1-SII(+) and narrower for the InsP3R1-SII(-) and InsP3R1-opt; 4) the apparent affinity for ATP is sixfold lower for the InsP3R1-SII(-) (1.4 mM) and 20-fold lower for the InsP3R1-opt (5.3 mM) than for the InsP3R1-SII(+) (0.24 mM); 5) the InsP3R1-SII(-) is approximately twofold more active than the InsP3R1-SII(+) in the absence of ATP. Obtained results provide novel information about the molecular determinants of the InsP3R1 function.

UR - http://www.scopus.com/inward/record.url?scp=0036217391&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036217391&partnerID=8YFLogxK

M3 - Article

C2 - 11916857

AN - SCOPUS:0036217391

VL - 82

SP - 1995

EP - 2004

JO - Biophysical Journal

JF - Biophysical Journal

SN - 0006-3495

IS - 4

ER -