Effects of tetraphenylboron-induced increase in inner surface charge on Ca2+ release from sarcoplasmic reticulum.
We studied the relationship between surface charge and release of Ca2+ in the heavy sarcoplasmic reticulum (SR) of skeletal muscle. The inner and outer surface potentials and charge densities of the membrane treated with a lipophilic anion, tetraphenylboron (TPB-), were measured using 1-anilino-8-naphthalene-sulfonate fluorescence. Ca2+ was loaded passively or actively by the SR. Ca2+ release was estimated by the fluorescence of chlortetracycline, and protein conformational change was monitored by use of the sulfhydryl group fluorescent probe, N-(7-dimethylamino-4-methyl-3-coumarinyl) maleimide (DACM). Treatment of Ca2(+)-loaded SR vesicles with micromolar TPB- dose-dependently increased the local fixed negative charge on the inner surface, and changed the DACM fluorescence intensity in parallel with the Ca2+ release. The changes in surface charge and in DACM fluorescence intensity did not originate from the Ca2+ flux. A lipophilic cation, tetraphenylarsonium (TPA+), screened the negative inner surface charge which was increased by TPB-, and inhibited both TPB(-)-induced change in DACM fluorescence intensity and Ca2+ release. Millimolar Mg2+ reduced degrees of TPB(-)-induced Ca2+ release from the SR and of TPB(-)-induced contraction in mechanically skinned fibers. Mg2+ did not inhibit the increase in the negative inner surface charge and DACM fluorescence intensity produced by TPB-. Thus, the local increase in negative charge on the SR inner membrane leaflet seems to be causally related to the Ca2+ release. Mg2+ and TPA+ are suggested to inhibit TPB(-)-induced Ca2+ release by different mechanisms.
Liu
G H
GH
Department of Physiology, Nagoya City University Medical School, Japan.
Oba
T
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D000818:Animals; D001152:Arsenicals / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D004563:Electrochemistry; D005456:Fluorescent Dyes; D066298:In Vitro Techniques; D007425:Intracellular Membranes / Q000378:metabolism; D008274:Magnesium / Q000494:pharmacology; D008301:Maleimides; D008564:Membrane Potentials / Q000187:drug effects; D011892:Rana catesbeiana; D012519:Sarcoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism*; D013775:Tetraphenylborate / Q000494:pharmacology
D001152:Arsenicals; D005456:Fluorescent Dyes; D008301:Maleimides; C009439:tetraphenylarsonium; D013775:Tetraphenylborate; C019347:N-(7-dimethylamino-4-methylcoumarinyl)maleimide; D008274:Magnesium; D002118:Calcium