ABCMdb

PMID: 11741811

Cobb BR, Ruiz F, King CM, Fortenberry J, Greer H, Kovacs T, Sorscher EJ, Clancy JP
A(2) adenosine receptors regulate CFTR through PKA and PLA(2).
Am J Physiol Lung Cell Mol Physiol. 2002 Jan;282(1):L12-25., [PubMed]

Sentences

No. Mutations Sentence Comment

17 ABCC7 p.Gly551Asp Ado also activated G551D CFTR-dependent halide efflux when combined with arachidonic acid and phosphodiesterase inhibition. Login to comment 51 ABCC7 p.Gly551Asp Wild-type (WT), ⌬F508, or G551D CFTR under control of the T7 promoter in the pTM-1 vector was then introduced into the cells in complex with N-[1-(2,3-dioleoyloxy)propyl]- N,N,N-trimethylammonium (DOTAP)-propane-1,2-dioleoyl- sn-glycero-3-phosphoethanolamine (DOPE; 20 ␮g DOTAP-DOPE and 5 ␮g pTM-1 CFTR/5 ϫ 105 cells) for 4 h. Login to comment 53 ABCC7 p.Gly551Asp Cells were then washed in PBS, returned to DMEM plus 10% FBS, and studied 18-24 h postinfection (for WT CFTR and G551D CFTR) or after being grown at 29°C for 48 h (⌬F508 CFTR). Login to comment 55 ABCC7 p.Gly551Asp To study CFTR activation, we measured halide efflux in COS-7 cells transiently expressing WT, ⌬F508, or G551D CFTR or in Calu-3 cells with the halide-quenched dye 6-methoxy-N-(3-sulfopropyl)quinolinium (SPQ; Molecular Probes, Eugene, OR) as previously described (16, 17). Login to comment 108 ABCC7 p.Ser489* The cftr(-/-) mice carried two copies of the human cftr cDNA, which contains a stop codon at position 489 (S489X). Login to comment 243 ABCC7 p.Gly551Asp A2B-ARs activate ⌬F508 CFTR and G551D CFTR. Login to comment 245 ABCC7 p.Gly551Asp To determine whether A2B receptor signaling could be used to improve the activity of common disease-causing CFTR mutations, we transiently expressed ⌬F508 CFTR (class II mutation) and G551D CFTR (class III mutation) in COS-7 cells. Login to comment 266 ABCC7 p.Gly551Asp In Fig. 11, COS-7 cells expressing G551D CFTR were stimulated with forskolin, Ado, or AA. Login to comment 269 ABCC7 p.Gly551Asp ABCC7 p.Gly551Asp Two nonspecific PDE inhibitors (papaverine and theophylline) were studied at 200 ␮M. PDE inhibitors such as milrinone have been evaluated in clinical trials for their ability to activate Cl-conductance in normal subjects and in CF patients carrying the G551D CFTR mutation (68). Login to comment 270 ABCC7 p.Gly551Asp At the concentrations used, none of the PDE inhibitors alone consistently elevated cAMP in COS-7 cells (10-min exposure; data not shown), and none of the stimuli (including Ado) activated G551D CFTR-specific halide efflux (Fig. 11A). Login to comment 272 ABCC7 p.Gly551Asp In Fig. 11B, G551D CFTR-expressing COS-7 cells were ex- Fig. 6. Login to comment 330 ABCC7 p.Gly551Asp Finally, we showed that the two most common disease-associated CFTR mutations can be activated by A2B receptors alone (⌬F508 CFTR) or by using messenger pathways stimulated by A2B receptors (G551D CFTR). Login to comment 355 ABCC7 p.Gly551Asp Activation of halide efflux in G551D CFTR-expressing cells. Login to comment 356 ABCC7 p.Gly551Asp A: G551D CFTR-expressing cells compared with wild-type (WT) CFTR-expressing cells. Login to comment 357 ABCC7 p.Gly551Asp COS-7 cells transiently expressing WT or G551D CFTR were studied with 6-methoxy-N-(3-sulfopropyl)quinolinium (SPQ) as described in METHODS; n 40 cells/condition. Login to comment 358 ABCC7 p.Gly551Asp G551D CFTR-expressing cells failed to activate halide efflux when stimulated with Ado (200 ␮M) ϩ papaverine (PAP; 200 ␮M), Ado ϩ rolipram (ROL; 20 ␮M) in combination, or Forsk (20 ␮M; arrow). Login to comment 360 ABCC7 p.Gly551Asp B: G551D CFTR-expressing COS-7 cells stimulated with Ado (200 ␮M) and AA (100 ␮M) combined with a series of phosphodiesterase inhibitors (PDEis). Login to comment 385 ABCC7 p.Gly551Asp The effect would need to be quite dramatic, however, because G551D CFTR, which has previously been shown to be poorly responsive to powerful phosphorylating stimuli, appears to be activated by costimulation with AA (Fig. 11B, Refs. Login to comment