Functional characterization of mammalian inositol 1,4,5-trisphosphate receptor isoforms

Huiping Tu, Zhengnan Wang, Elena Nosyreva, Humbert De Smedt, Ilya Bezprozvanny

Research output: Contribution to journalArticle

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Abstract

Inositol 1,4,5-trisphosphate receptors (InsP3R) play a key role in intracellular calcium (Ca2+) signaling. Three mammalian InsP 3R isoforms-InsP3R type 1 (InsP3R1), InsP 3R type 2 (InsP3R2), and InsP3R type 3 (InsP3R3) are expressed in mammals, but the functional differences between the three mammalian InsP3R isoforms are poorly understood. Here we compared single-channel behavior of the recombinant rat InsP 3R1, InsP3R2, and InsP3R3 expressed in Sf9 cells, reconstituted into planar lipid bilayers and recorded with 50 mM Ba 2+ as a current carrier. We found that: 1), for all three mammalian InsP3R isoforms the size of the unitary current is 1.9 pA and single-channel conductance is 74-80 pS; 2), in optimal recording conditions the maximal single-channel open probability for all three mammalian InsP 3R isoforms is in the range 30-40%; 3), in optimal recording conditions the mean open dwell time for all three mammalian InsP3R isoforms is 7-8 ms, the mean closed dwell time is ∼10 ms; 4), InsP 3R2 has the highest apparent affinity for InsP3 (0.10 μ), followed by InsP3R1 (0.27 μM), and then by InsP 3R3 (0.40 μM); 5), InsP3R1 has a high-affinity (0.13 mM) ATP modulatory site, InsP3R2 gating is ATP independent, and InsP3R3 has a low-affinity (2 mM) ATP modulatory site; 6), ATP modulates InsP3R1 gating in a noncooperative manner (nHill = 1.3); 7), ATP modulates InsP3R3 gating in a highly cooperative manner (nHill = 4.1). Obtained results provide novel information about functional properties of mammalian InsP3R isoforms.

Original languageEnglish (US)
Pages (from-to)1046-1055
Number of pages10
JournalBiophysical Journal
Volume88
Issue number2
DOIs
StatePublished - Feb 2005

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Inositol 1,4,5-Trisphosphate Receptors
Protein Isoforms
Adenosine Triphosphate
Sf9 Cells
Calcium Signaling
Lipid Bilayers
Mammals

ASJC Scopus subject areas

  • Biophysics

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Functional characterization of mammalian inositol 1,4,5-trisphosphate receptor isoforms. / Tu, Huiping; Wang, Zhengnan; Nosyreva, Elena; De Smedt, Humbert; Bezprozvanny, Ilya.

In: Biophysical Journal, Vol. 88, No. 2, 02.2005, p. 1046-1055.

Research output: Contribution to journalArticle

Tu, Huiping ; Wang, Zhengnan ; Nosyreva, Elena ; De Smedt, Humbert ; Bezprozvanny, Ilya. / Functional characterization of mammalian inositol 1,4,5-trisphosphate receptor isoforms. In: Biophysical Journal. 2005 ; Vol. 88, No. 2. pp. 1046-1055.
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abstract = "Inositol 1,4,5-trisphosphate receptors (InsP3R) play a key role in intracellular calcium (Ca2+) signaling. Three mammalian InsP 3R isoforms-InsP3R type 1 (InsP3R1), InsP 3R type 2 (InsP3R2), and InsP3R type 3 (InsP3R3) are expressed in mammals, but the functional differences between the three mammalian InsP3R isoforms are poorly understood. Here we compared single-channel behavior of the recombinant rat InsP 3R1, InsP3R2, and InsP3R3 expressed in Sf9 cells, reconstituted into planar lipid bilayers and recorded with 50 mM Ba 2+ as a current carrier. We found that: 1), for all three mammalian InsP3R isoforms the size of the unitary current is 1.9 pA and single-channel conductance is 74-80 pS; 2), in optimal recording conditions the maximal single-channel open probability for all three mammalian InsP 3R isoforms is in the range 30-40{\%}; 3), in optimal recording conditions the mean open dwell time for all three mammalian InsP3R isoforms is 7-8 ms, the mean closed dwell time is ∼10 ms; 4), InsP 3R2 has the highest apparent affinity for InsP3 (0.10 μ), followed by InsP3R1 (0.27 μM), and then by InsP 3R3 (0.40 μM); 5), InsP3R1 has a high-affinity (0.13 mM) ATP modulatory site, InsP3R2 gating is ATP independent, and InsP3R3 has a low-affinity (2 mM) ATP modulatory site; 6), ATP modulates InsP3R1 gating in a noncooperative manner (nHill = 1.3); 7), ATP modulates InsP3R3 gating in a highly cooperative manner (nHill = 4.1). Obtained results provide novel information about functional properties of mammalian InsP3R isoforms.",
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