Modulation of BK channel calcium affinity by differential phosphorylation in developing ovine basilar artery myocytes.
Large-conductance Ca2+-sensitive K+ (BK) channel activity is greater in basilar artery smooth muscle cells (SMCs) of the fetus than the adult, and this increased activity is associated with a lower BK channel Ca2+ set point (Ca0). Associated PKG activity is three times greater in BK channels from fetal than adult myocytes, whereas associated PKA activity is three times greater in channels from adult than fetal myocytes. We hypothesized that the change in Ca0 during development results from different levels of channel phosphorylation. In inside-out membrane patch preparations of basilar artery SMCs from adult and fetal sheep, we measured BK channel activity in four states of phosphorylation: native, dephosphorylated, PKA phosphorylated, and PKG phosphorylated. BK channels from adult and fetus exhibited similar voltage-activation curves, Ca0 values, and Ca2+ dissociation constants (Kd) for the dephosphorylated, PKA phosphorylated, and PKG phosphorylated states. However, voltage-activation curves of native fetal BK channels shifted significantly to the left of those of the adult, with Ca0 and Kd values half those of the adult. For the two age groups at each of the phosphorylation states, Ca0 and Kd produced linear relations when plotted against voltage at half-maximal channel activation. We conclude that the Ca0 and Kd values of the BK channel can be modulated by differential channel phosphorylation. Lower Ca0 and Kd values in BK channels of fetal myocytes can be explained by a greater extent of channel phosphorylation of fetal than adult myocytes.
Lin
Mike T
MT
Center for Perinatal Biology, Loma Linda University School of Medicine, Loma Linda, CA 92350, USA.
Hessinger
David A
DA
Pearce
William J
WJ
Longo
Lawrence D
LD
D000375:Aging / Q000502:physiology; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001488:Basilar Artery / Q000166:cytology / Q000254:growth & development* / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D002469:Cell Separation; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism; D005260:Female; D005333:Fetus / Q000378:metabolism; D007202:Indicators and Reagents; D007700:Kinetics; D051036:Large-Conductance Calcium-Activated Potassium Channels / Q000502:physiology*; D008564:Membrane Potentials / Q000502:physiology; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D010766:Phosphorylation; D011247:Pregnancy; D012756:Sheep
D007202:Indicators and Reagents; D051036:Large-Conductance Calcium-Activated Potassium Channels; D017868:Cyclic AMP-Dependent Protein Kinases; D017869:Cyclic GMP-Dependent Protein Kinases; D000469:Alkaline Phosphatase; D002118:Calcium