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Chloride concentration affects Kv channel voltage-gating kinetics: Importance of experimental anion concentrations.
- Source :
-
Brain research bulletin [Brain Res Bull] 2005 Sep 30; Vol. 67 (1-2), pp. 142-6. - Publication Year :
- 2005
-
Abstract
- Chloride concentration has been shown to have a dramatic impact on protein folding and subsequent tertiary conformation [K.D. Collins, Ions from the Hofmeister series and osmolytes: effects on proteins in solution and in the crystallization process, Methods 34 (2004) 300-311; I. Jelesarov, E. Durr, R.M. Thomas, H.R. Bosshard, Salt effects on hydrophobic interaction and charge screening in the folding of a negatively charged peptide to a coiled coil (leucine zipper), Biochemistry 37 (1998) 7539-7550]. As it is known that Kv channel gating is linked to the stability of the cytoplasmic T1 multimerization domain conformation [D.L. Minor, Y.F. Lin, B.C. Mobley, A. Avelar, Y.N. Jan, L.Y. Jan, J.M. Berger, The polar T1 interface is linked to conformational changes that open the voltage-gated potassium channel, Cell 102 (2000) 657-670; B.A. Yi, D.L. Minor Jr., Y.F. Lin, Y.N. Jan, L.Y. Jan, Controlling potassium channel activities: interplay between the membrane and intracellular factors, Proc. Natl. Acad. Sci. U.S.A. 98 (2001) 11016-11023] and that intracellular chloride concentration has been linked to Kv channel kinetics [L.K. Bekar, W. Walz, Intracellular chloride modulates A-type potassium currents in astrocytes, Glia 39 (2002) 207-216; W.B. Thoreson, S.L. Stella, Anion modulation of calcium current voltage dependence and amplitude in salamander rods, Biochim. Biophys. Acta 1464 (2000) 142-150], the objective of the present study was to address how chloride concentration changes affect Kv channel kinetics more closely in an isolated expression system. Initially, no significant chloride concentration-dependent effects on channel steady-state gating kinetics were observed. Only after disruption of the cytoskeleton with cytochalasin-D did we see significant chloride concentration-dependent shifts in gating kinetics. This suggests that the shift in gating kinetics is mediated through effects of intracellular chloride concentration on cytoplasmic domain tertiary conformation as cytoskeletal interaction appears to mask the effect. Furthermore, as cytoskeletal disruption only impacts channel gating kinetics at low physiological intracellular chloride concentrations, these studies highlight the importance of paying close attention to anion concentrations used under experimental conditions.
- Subjects :
- Cell Line
Cell Membrane drug effects
Chlorides pharmacology
Cytoskeleton drug effects
Cytoskeleton physiology
Humans
Intracellular Fluid metabolism
Ion Channel Gating drug effects
Kinetics
Membrane Potentials drug effects
Membrane Potentials physiology
Patch-Clamp Techniques
Potassium Channels, Voltage-Gated drug effects
Cell Membrane metabolism
Chlorides metabolism
Ion Channel Gating physiology
Potassium Channels, Voltage-Gated metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 0361-9230
- Volume :
- 67
- Issue :
- 1-2
- Database :
- MEDLINE
- Journal :
- Brain research bulletin
- Publication Type :
- Academic Journal
- Accession number :
- 16140173
- Full Text :
- https://doi.org/10.1016/j.brainresbull.2005.06.023