PKA holoenzyme is functionally coupled to CFTR by AKAPs

P. Huang, K. Trotter, R. C. Boucher, S. L. Milgram, M. J. Stutts
2000 American Journal of Physiology - Cell Physiology  
Cystic fibrosis transmembrane regulator (CFTR) is reported to be preferentially regulated by membranebound protein kinase A (PKAII). We tested for close physical and functional association of PKA with CFTR in inside-out membrane patches excised from Calu-3 cells. In the presence of MgATP, 8-(4-chlorophenylthio)adenosine 3Ј,5Ј-cyclic monophosphate (CPT-cAMP) increased the product of CFTR channel number and open probability (from 0.36 Ϯ 0.12 to 1.23 Ϯ 0.57, n ϭ 20, P Ͻ 0.0025), and this
more » ... and this stimulation was abolished by PKI. Thus Calu-3 membrane isolated from cells retains PKA holoenzyme that is functionally coupled to CFTR. PKAII is anchored at specific subcellular sites by A kinase anchoring proteins (AKAPs). Exposure of excised patches to HT-31, a peptide that disrupts the association of PKAII and AKAPs, prevented CPT-cAMP stimulation of CFTR. Therefore, PKA holoenzyme in isolated membrane patches is bound to AKAPs. In whole cell voltage-clamp studies, intracellular dialysis of Calu-3 cells with HT-31 blocked the activation of CFTR by extracellular adenosine. These results suggest that AKAPs mediate PKA compartmentalization with CFTR and are required for activation of CFTR by physiological regulators.
doi:10.1152/ajpcell.2000.278.2.c417 pmid:10666038 fatcat:g44vszz4tjd5rhh4z6e4yqgbsm