pH-dependent interactions of Cd2+ and a carboxylate blocker with the rat C1C-1 chloride channel and its R304E mutant in the Sf-9 insect cell line

The Journal of Physiology
G Y RychkovM L Roberts

Abstract

1. Gating of the skeletal muscle chloride channel (ClC-1) is sensitive to extracellular pH. In this study, whole-cell recording of currents from wild-type (WT) ClC-1 and a mutant, R304E, expressed in the Sf-9 insect cell line was used to investigate further the nature of the pH-sensitive residues. 2. Extracellular Cd2+ produced a concentration-dependent block of WT ClC-1 with an IC50 of 1.0 +/- 0.1 mM and a Hill coefficient of 2.0 +/- 0.3. This block was sensitive to external pH, reducing at low pH, with an apparent pKa of 6.8 +/- 0.1 and a Hill coefficient for proton binding of 3.0 +/- 0.3. Anthracene-9-carboxylate (A-9-C) block of WT ClC-1 was also pH sensitive, increasing at low pH, with an apparent pKa of 6.4 +/- 0.1 and a Hill coefficient for proton binding of 1.0 +/- 0.2. 3. Compared with WT ClC-1, R304E had a lower affinity for Cd2+ (IC50, 3.0 +/- 0.3 mM) but it had a similar Hill coefficient for transition metal ion binding. The Hill coefficient for proton binding to the Cd2+ binding site was reduced to 1.4 +/- 0.3. In contrast, the A-9-C binding site in R304E showed the same pH sensitivity and affinity for the blocker as that seen in WT ClC-1. 4. ClC-1 has at least two binding sites for Cd2+, each of which has at least...Continue Reading

References

Jun 1, 1977·The Journal of General Physiology·P T Palade, R L Barchi
Apr 1, 1987·Physiological Reviews·A H Bretag
Dec 1, 1971·The Journal of Physiology·S H Bryant, A Morales-Aguilera
Apr 1, 1967·The Journal of Physiology·O F Hutter, A E Warner
Apr 1, 1967·The Journal of Physiology·O F Hutter, A E Warner
Jul 1, 1983·The Journal of General Physiology·W Hanke, C Miller
Nov 4, 1993·Nature·J A TabcharaniJ W Hanrahan
Apr 21, 1995·The Journal of Biological Chemistry·C A GurnettK P Campbell
Jan 1, 1995·The Journal of Physiology·T J JentschB Schwappach
Dec 1, 1996·The Journal of Physiology·G Y RychkovA H Bretag

❮ Previous
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Citations

Feb 28, 1974·Research in Experimental Medicine. Zeitschrift Für Die Gesamte Experimentelle Medizin Einschliesslich Experimenteller Chirurgie·R Bodem, E Kuhn
Apr 27, 1999·British Journal of Pharmacology·E C AromatarisM L Roberts
Jun 20, 1998·The Journal of General Physiology·G Y RychkovA H Bretag
Dec 2, 1998·The Journal of General Physiology·T Y Chen
Jun 29, 2001·The Journal of General Physiology·M F Chen, T Y Chen
Jul 2, 2008·The Journal of General Physiology·Giovanni ZifarelliMichael Pusch
Nov 17, 2010·The Journal of General Physiology·John D LueckRobert T Dirksen
Aug 17, 2001·The Journal of Physiology·B BennettsG Y Rychkov
Feb 16, 2005·Annual Review of Physiology·Tsung-Yu Chen
Jun 22, 2000·Proceedings of the National Academy of Sciences of the United States of America·T K RostovtsevaS M Bezrukov
Dec 2, 2015·The Journal of General Physiology·Yawei YuTsung-Yu Chen
Mar 21, 1998·The Journal of Biological Chemistry·D H HryciwA H Bretag
Aug 26, 1998·Nature·C FahlkeA L George
May 31, 2018·Physiological Reviews·Thomas J Jentsch, Michael Pusch
Sep 2, 2004·Proteins·Jian YinThomas L Beck
Oct 18, 2006·Clinical and Experimental Pharmacology & Physiology·Edoardo C Aromataris, Grigori Y Rychkov
Apr 9, 2008·Physiological Reviews·Tsung-Yu Chen, Tzyh-Chang Hwang
May 10, 2001·The Journal of Biological Chemistry·M E HartnessP J Kemp
May 17, 2002·The Journal of Physiology·Jorge ArreolaJames E Melvin
Mar 29, 2003·Physiological Reviews·Thomas E Decoursey
Oct 29, 2000·American Journal of Physiology. Gastrointestinal and Liver Physiology·E K O'DonnellB D Schultz
Jul 9, 2005·The Journal of Physiology·Michael D DuffieldMichael L Roberts
Oct 3, 1999·Physiological Reviews·F Lehmann-Horn, K Jurkat-Rott

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