The Journal of General Physiology
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Published 1 March 2000. doi:10.1085/jgp.115.3.241
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© The Rockefeller University Press, 0022-1295/2000//241/ $5.00
Journal of General Physiology, Volume 115, Number 3, 2000


Original Article

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

Don-On Daniel Maka, Sean McBridea, Viswanathan Raghurama, Yun Yuea, Suresh K. Josephc, and J. Kevin Fosketta,b

a From the Department of Physiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104
b From the Institute for Human Gene Therapy, University of Pennsylvania, Philadelphia, Pennsylvania 19104
c Department of Pathology, Thomas Jefferson University School of Medicine, Philadelphia, Pennsylvania 19107
Department of Physiology, University of Pennsylvania, B400 Richards Building, Philadelphia, PA 19104.215-573-6808

foskett{at}mail.med.upenn.edu

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.

Key Words: calcium • calcium release channel • electrophysiology • expression • Xenopus


© 2000 The Rockefeller University Press


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D.-O. D. Mak, S. McBride, and J. K. Foskett
Regulation by Ca2+ and Inositol 1,4,5-Trisphosphate (Insp3) of Single Recombinant Type 3 Insp3 Receptor Channels: Ca2+ Activation Uniquely Distinguishes Types 1 and 3 Insp3 Receptors
J. Gen. Physiol., May 1, 2001; 117(5): 435 - 446.
[Abstract] [Full Text] [PDF]


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D.-O. D. Mak, S. McBride, and J. K. Foskett
Atp Regulation of Recombinant Type 3 Inositol 1,4,5-Trisphosphate Receptor Gating
J. Gen. Physiol., May 1, 2001; 117(5): 447 - 456.
[Abstract] [Full Text] [PDF]



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