ABCMdb

PMID: 22999299

Tucker TA, Fortenberry JA, Zsembery A, Du M, Bedwell DM, Schwiebert LM, Schwiebert EM
The DeltaF508-CFTR mutation inhibits wild-type CFTR processing and function when co-expressed in human airway epithelia and in mouse nasal mucosa.
BMC Physiol. 2012 Sep 24;12(1):12., [PubMed]

Sentences

No. Mutations Sentence Comment

74 ABCC7 p.Trp1282* The IB3-1 cell line (derived from a CF human bronchus expressing the ƊF508 and W1282X mutant forms of CFTR) was grown in LHC-8 media without gentamycin (Biofluids) supplemented with 5% heat-inactivated fetal bovine serum, 6 ml of penicillin-streptomycin 100x solution (penicillin 100 U/ml and streptomycin 100 bc;g/mg final), 6 ml of 200 mM L-glutamine 100X solution (2 mM final), and 2 ml of fungizone solution (amphotericin B, 1 ug/ml final). Login to comment 79 ABCC7 p.Trp1282* The IB3-1 cell line (derived from a CF human bronchus expressing the ΔF508 and W1282X mutant forms of CFTR) was grown in LHC-8 media without gentamycin (Biofluids) supplemented with 5% heat-inactivated fetal bovine serum, 6 ml of penicillin-streptomycin 100x solution (penicillin 100U/ml and streptomycin 100 μg/ mg final), 6 ml of 200 mM L-glutamine 100X solution (2 mM final), and 2 ml of fungizone solution (amphotericin B, 1ug/ml final). Login to comment 82 ABCC7 p.Gly551Asp ABCC7 p.Gly551Asp For G551D-CFTR experiments, the same amounts of G551D-CFTR bearing plasmid were used as a substitute for ƊF-CFTR. Login to comment 87 ABCC7 p.Gly551Asp ABCC7 p.Gly551Asp For G551D-CFTR experiments, the same amounts of G551D-CFTR bearing plasmid were used as a substitute for ΔF-CFTR. Login to comment 222 ABCC7 p.Gly551Asp We wished to take addition steps to provide specificity and exclusivity for this macromolecular complex and PDZ motif-driven ƊF-CFTR/WT-CFTR inhibitory interaction, we expressed increasing amounts of G551D-CFTR with a fixed amount of WT-CFTR. Login to comment 224 ABCC7 p.Gly551Asp It is important to underscore the fact that G551D-CFTR is not an ER retention mutant but rather is processed normally to the plasma membrane. Login to comment 225 ABCC7 p.Gly551Asp Rather, G551D-CFTR is a dysfunctional Cl-channel because ATP binding and gating to its NBDs is impaired. Login to comment 226 ABCC7 p.Gly551Asp ABCC7 p.Gly551Asp This is why the CFTR potentiator drug, VX-770 (ivacaftor, Kalydeco) [54] is markedly effective in G551D-CFTR patients, while the CF corrector drug, VX-809, corrects/rescues ƊF-CFTR from ER quality control and is without effect on G551D-CFTR patients [55,56]. Login to comment 233 ABCC7 p.Gly551Asp We wished to take addition steps to provide specificity and exclusivity for this macromolecular complex and PDZ motif-driven ΔF-CFTR/WT-CFTR inhibitory interaction, we expressed increasing amounts of G551D-CFTR with a fixed amount of WT-CFTR. Login to comment 235 ABCC7 p.Gly551Asp It is important to underscore the fact that G551D-CFTR is not an ER retention mutant but rather is processed normally to the plasma membrane. Login to comment 236 ABCC7 p.Gly551Asp Rather, G551D-CFTR is a dysfunctional Cl- channel because ATP binding and gating to its NBDs is impaired. Login to comment 237 ABCC7 p.Gly551Asp ABCC7 p.Gly551Asp This is why the CFTR potentiator drug, VX-770 (ivacaftor, Kalydeco) [54] is markedly effective in G551D-CFTR patients, while the CF corrector drug, VX-809, corrects/rescues ΔF-CFTR from ER quality control and is without effect on G551D-CFTR patients [55,56]. Login to comment 244 ABCC7 p.Gly551Asp Similar co-expression biochemical experiments with G551D-CFTR and WT-CFTR. Login to comment 260 ABCC7 p.Gly551Asp 50-70 kDa Gly. P2X4 WT: F 1:1 WT: F 1:2 WT: F 1:4 B 1:4 1:2 1:1 G551D-CFTR WT-CFTR EV Band B Band C WT:G551D A Figure 2 Confirming the specificity of the dominant negative-like effect of ƊF-CFTR on WT-CFTR in native IB3-1 CF bronchial epithelial cells. Login to comment 262 ABCC7 p.Gly551Asp Similar co-expression biochemical experiments with G551D-CFTR and WT-CFTR. Login to comment 586 ABCC7 p.Gly551Asp Accurso FJ, et al: Effect of VX-770 in persons with cystic fibrosis and the G551D-CFTR mutation. Login to comment