TY - JOUR
T1 - Modulation of the Ca2+-activated large conductance K+ channel by intracellular pH in human renal proximal tubule cells
AU - Hirano, Junko
AU - Nakamura, Kazuyoshi
AU - Itazawa, Shun Ichi
AU - Sohma, Yoshiro
AU - Kubota, Takahiro
AU - Kubokawa, Manabu
PY - 2002/6
Y1 - 2002/6
N2 - The Ca2+-activated and voltage-sensitive large conductance K- channel (BK channel) with a slope conductance of about 300 pS is present in the surface membrane of cultured human renal proximal tubule epithelial cells (RPTECs). In this study we examined the effects of cytoplasmic pH (pHi) on activity and gating kinetics of the BK channel by using the inside-out configuration of the patch-clamp technique. At a constant cytoplasmic Ca2- concentration ([Ca2-]i), membrane depolarization raised channel open probability (Po), and lowering pHi shifted the Po-membrane potential (Vm) relationship to the positive voltage direction. However, the value of the gating charge was not affected by changes in pHi, suggesting that the effects of pHi on Po were not due to an alternation of the voltage sensitivity. At constant Vm, lowering pHi suppressed the [Ca2+]i-dependent channel activation and shifted the Po-[Ca2+]i relationship in the direction of higher [Ca2-]i with a reduction of maximal Po. Furthermore, both the mean open and mean closed times of the BK channels at pHi 6.3 in the presence of 10-4M [Ca2-]i were shorter than those at pHi 7.3 in the presence of 10-5 M [Ca2+]i, even though these two different conditions gave a similar Po. The data indicate that cytoplasmic H- suppresses Po of the BK channel in RPTECs, which involves the mechanism independent of Ca2+ activation. Our preliminary kinetic analysis also supported this notion.
AB - The Ca2+-activated and voltage-sensitive large conductance K- channel (BK channel) with a slope conductance of about 300 pS is present in the surface membrane of cultured human renal proximal tubule epithelial cells (RPTECs). In this study we examined the effects of cytoplasmic pH (pHi) on activity and gating kinetics of the BK channel by using the inside-out configuration of the patch-clamp technique. At a constant cytoplasmic Ca2- concentration ([Ca2-]i), membrane depolarization raised channel open probability (Po), and lowering pHi shifted the Po-membrane potential (Vm) relationship to the positive voltage direction. However, the value of the gating charge was not affected by changes in pHi, suggesting that the effects of pHi on Po were not due to an alternation of the voltage sensitivity. At constant Vm, lowering pHi suppressed the [Ca2+]i-dependent channel activation and shifted the Po-[Ca2+]i relationship in the direction of higher [Ca2-]i with a reduction of maximal Po. Furthermore, both the mean open and mean closed times of the BK channels at pHi 6.3 in the presence of 10-4M [Ca2-]i were shorter than those at pHi 7.3 in the presence of 10-5 M [Ca2+]i, even though these two different conditions gave a similar Po. The data indicate that cytoplasmic H- suppresses Po of the BK channel in RPTECs, which involves the mechanism independent of Ca2+ activation. Our preliminary kinetic analysis also supported this notion.
KW - Channel activity
KW - Gating kinetics
KW - Intracellular pH
KW - Kidney
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U2 - 10.2170/jjphysiol.52.267
DO - 10.2170/jjphysiol.52.267
M3 - Article
C2 - 12230803
AN - SCOPUS:0036592067
SN - 1880-6546
VL - 52
SP - 267
EP - 276
JO - Journal of Physiological Sciences
JF - Journal of Physiological Sciences
IS - 3
ER -