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The major cystic fibrosis causing mutation exhibits defective propensity for phosphorylation
- Source :
- PROTEOMICS. 15:447-461
- Publication Year :
- 2014
- Publisher :
- Wiley, 2014.
-
Abstract
- The major cystic fibrosis causing mutation, F508del-CFTR (where CFTR is cystic fibrosis transmembrane conductance regulator), impairs biosynthetic maturation of the CFTR protein, limiting its expression as a phosphorylation-dependent channel on the cell surface. The maturation defect can be partially rescued by low-temperature (27°C) cell culture conditions or small-molecule corrector compounds. Following its partial rescue, the open probability of F508del-CFTR is enhanced by the potentiator compound, VX-770. However, the channel activity of rescued F508del-CFTR remains less than that of the Wt-CFTR protein in the presence of VX-770. In this study, we asked if there are allosteric effects of F508del on the phosphorylation-regulated R domain. To identify defects in the R domain, we compared the phosphorylation status at protein kinase A sites in the R domain of Wt and F508del-CFTR. Here we show that phosphorylation of Ser-660, quantified by SRM-MS, is reduced in F508del-CFTR. Although the generation of a phosphomimic at this site (substituting aspartic acid for serine) did not modify the maturation defect, it did enhance F508del-CFTR channel function after pharmacological rescue with corrector VX-809, and treatment with the potentiator, VX-770. These findings support the concept that defective phosphorylation of F508del-CFTR partially accounts for its altered channel activity at the cell surface.
- Subjects :
- congenital, hereditary, and neonatal diseases and abnormalities
Cystic Fibrosis
Molecular Sequence Data
Allosteric regulation
Cystic Fibrosis Transmembrane Conductance Regulator
Biology
medicine.disease_cause
Biochemistry
Cystic fibrosis
Cell Line
Serine
Cricetinae
medicine
Animals
Humans
Amino Acid Sequence
Phosphorylation
Protein kinase A
Molecular Biology
Sequence Deletion
Mutation
respiratory system
Potentiator
medicine.disease
Cyclic AMP-Dependent Protein Kinases
Molecular biology
Cystic fibrosis transmembrane conductance regulator
Protein Structure, Tertiary
respiratory tract diseases
HEK293 Cells
biology.protein
Subjects
Details
- ISSN :
- 16159853
- Volume :
- 15
- Database :
- OpenAIRE
- Journal :
- PROTEOMICS
- Accession number :
- edsair.doi.dedup.....ee59295b1785e79aa8adc91d44c1de47