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J. Biol. Chem., Vol. 276, Issue 2, 1063-1070, January 12, 2001
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From the Laboratory of Signal Transduction, NIEHS, National
Institutes of Health,
Research Triangle Park, North Carolina 27709
The regulation of store-operated,
calcium-selective channels in the plasma membrane of rat basophilic
leukemia cells (RBL-2H3 m1), an immortalized mucosal mast cell line,
was studied at the single-channel level with the patch clamp technique
by removing divalent cations from both sides of the membrane. The
activity of the single channels in excised patches could be modulated
by Ca2+, Mg2+, and pH. The maximal
activation of these channels by divalent cation-free conditions
occurred independently of depletion of intracellular Ca2+
stores, whether in excised patches or in whole cell mode. Yet, a number
of points of evidence establish these single-channel openings as
amplified store-operated channel events. Specifically, (i) the single
channels are exquisitely sensitive to inhibition by intracellular
Ca2+, and (ii) both the store-operated current and the
single-channel openings are completely blocked by the capacitative
calcium entry blocker, 2-aminoethoxydiphenyl borane. In addition, in
Jurkat T cells single-channel openings with lower open probability have been observed in the whole cell mode with intracellular
Mg2+ present (Kerschbaum, H. H., and Cahalan, M. D. (1999) Science 283, 836-839), and in RBL-2H3 m1 cells a
current with similar properties is activated by store depletion.
Stable Activation of Single Ca2+ Release-activated
Ca2+ Channels in Divalent Cation-free Solutions*
,
*
The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed. Tel.: 919-541-3298;
Fax: 919-541-1898; E-mail: braun2@niehs.nih.gov.
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