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Am J Physiol Renal Physiol 285: F289-F294, 2003. First published April 1, 2003; doi:10.1152/ajprenal.00046.2003
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Role of h{beta}1 in activation of human mesangial BK channels by cGMP kinase

Patrick E. Kudlacek, Jennifer L. Pluznick, Rong Ma, Babu Padanilam, and Steven C. Sansom

Department of Physiology and Biophysics, University of Nebraska Medical Center, Omaha, Nebraska 68198-4575

Submitted 4 February 2003 ; accepted in final form 30 March 2003

In vascular smooth muscle and glomerular mesangial cells, relaxing agents such as nitric oxide and atrial natriuretic peptide activate large-conductance Ca2+-activated K+ channels (BK) via the cGMP kinase pathway. BK are composed of pore-forming {alpha}-subunits, encoded by the slopoke gene (Slo), and one of four cell-specific accessory {beta}-subunits (h{beta}1–4). We used patch-clamp analysis to determine the influence of h{beta}1, h{beta}2, and h{beta}4 on activation of human mesangial BK by cGMP kinase. We found that HEK 293 cells, coexpressing human (h) Slo{alpha} with either h{beta}1 or h{beta}2, contained single BK currents activated by db-cGMP in cell-attached patches. However, recombinant BK were not activated by db-cGMP when hSlo{alpha} was expressed alone or with h{beta}4. DNA-RNA hybridization revealed that mesangial cells contained mRNA for h{beta}1 but not h{beta}2 or h{beta}4. The BK response to db-cGMP was decreased when h{beta}1 antisense but not scrambled oligonucleotides were incorporated into mesangial cells. Western blot analysis showed that h{beta}1 antisense oligonucleotide inhibited the amount of h{beta}1-V5 fusion protein expressed in HEK 293 cells by ~50%. These results show that mesangial cells contain h{beta}1, a BK accessory protein, which confers activation of BK by cGMP kinase.

large-conductance calcium-activated potassium channels; maxi-potassium; human {beta}-subunit; antisense; patch clamp; guanosine 3',5'-cyclic monophosphate


Address for reprint requests and other correspondence: S. C. Sansom, Dept. of Physiology and Biophysics, 984575 Nebraska Medical Center, Omaha, NE 68198-4575 (E-mail: ssansom{at}unmc.edu).




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