ABCMdb

PMID: 24816489

Favia M, Mancini MT, Bezzerri V, Guerra L, Laselva O, Abbattiscianni AC, Debellis L, Reshkin SJ, Gambari R, Cabrini G, Casavola V
Trimethylangelicin promotes the functional rescue of mutant F508del CFTR protein in cystic fibrosis airway cells.
Am J Physiol Lung Cell Mol Physiol. 2014 Jul 1;307(1):L48-61. doi: 10.1152/ajplung.00305.2013. Epub 2014 May 9., [PubMed]

Sentences

No. Mutations Sentence Comment

46 ABCC7 p.Pro574His ABCC7 p.Pro574His Fischer rat thyroid (FRT) epithelial cells, stably coexpressing human F508del CFTR and the high-sensitivity halide-sensing green fluorescent analog yellow fluorescent protein (HS-YFP) YFP-H148Q/ I152L (FRT-F508del) or stably transfected with a plasmid coding for P574H-CFTR and carrying the resistance gene for zeocin (FRT-P574H) were a generous gift from Dr. L. J. Galietta (Gaslini Institute, Genoa, Italy). Login to comment 47 ABCC7 p.Pro574His FRT-F508del and FRT-P574H cells were grown in Coon`s modified Ham`s F-12 medium plus 10% FBS, L-glutamine, and penicillin/streptomycin at 37&#b0;C under 5% CO2. Login to comment 279 ABCC7 p.Pro574His ABCC7 p.Pro574His Last, to analyze if the correction effect of TMA was specific for the F508del mutation of CFTR, TMA corrector activity was tested on FRT cells stably expressing P574H-CFTR (FRT-P574H), a mutant of CFTR that, like F508del CFTR, is retained at the ER and can be rescued by incubation for 24 h at reduced temperature (42). Login to comment 280 ABCC7 p.Pro574His FRT-P574H cell monolayers were incubated for 24 h with 100 nM TMA or at 27&#b0;C, and the chloride efflux in response to FSK plus IBMX was measured as described in MATERIALS AND METHODS. Login to comment 287 ABCC7 p.Pro574His produce a significant correction in FRT-P574H cells [0.0005 afe; 0.0003 èc;(F/F0)/min, n afd; 4], with respect to the positive low-temperature rescue response [0.011 afe; 0.001 èc;(F/F0)/min, n afd; 5], suggesting that the TMA correction effect is specific for F508del mutation of CFTR. Login to comment 321 ABCC7 p.Pro574His Importantly, we found that TMA corrector activity is specific for the F508del mutation since TMA incubation fails to activate the temperature-sensitive mutant P574H-CFTR (42). Login to comment 333 ABCC7 p.Pro574His J. Zabner for providing CuFi-1 cells, Dr. L. J. Galietta for providing FRT cells stably coexpressing human F508del CFTR and high-sensitivity halide-sensing GFP analog YFP H148Q/ I152L and the P574H-CFTR FRT cells, and Dr. E. Ficker for providing the mutated constructs of hERG. Login to comment