Phosphorylation of the R domain by cAMP-dependent protein kinase regulates the CFTR chloride channel

Cheng, Shu‐Meng; Rich, Devra P.; Marshall, John; Gregory, Richard J.; Welsh, Michael J.; Smith, Adam E.

doi:10.1016/0962-8924(91)90010-7

Cited by 141 publications

(211 citation statements)

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“…Initial in vivo studies showed that four major sites-serines 660,737.795, and 81 3-were labeled after PKA simulation (Cheng et al 1991). However, mutation (Ser + Ala) of all these four sites did not eliminate CAMPdependent activation of CFTR (Chang et al.…”

Section: E T L H R E S L E G D a P V S W L E T K~f~t G E F G E K R K mentioning

confidence: 99%

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Identification of protein kinase A phosphorylation sites on NBD1 and R domains of CFTR using electrospray mass spectrometry with selective phosphate ion monitoring

et al. 1996

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HPLC-electrospray mass spectrometry was used to identify the phosphorylated sites on a bacterially expressed cystic fibrosis transmembrane conductance regulator (CFTR) fragment containing the first nucleotide binding domain (NBDI) and the regulatory domain (R). Tryptic digests of NBDI-R (CFTR residues 404-830) were analyzed after protein kinase A (PKA) treatment for all possible peptides and phosphopeptides (a total of 1 18 species) containing Ser residues within "high-probability'' PKA consensus sequences: R-R/K-X-SIT, R-X-X-SIT, and R-X-S/T. Three criteria were used to assign phosphorylated sites: ( I ) an 80-Da increase in the predicted average molecular weight of the tryptic peptides; ( 2 ) co-elution with the PO,-ion induced by stepped energy collision; and (3) the relative elution positions of the phosphorylated and unmodified peptides. Ser residues within the eight dibasic sites in the NBDl and R domains (positions 422, 660, 700, 712, 737, 768, 795, and 813) were phosphorylated, a pattern similar to that observed for full-length CFTR. The serine at position 753, which in CFTR is phosphorylated in vivo, was not phosphorylated. The remaining potential PKA sites, Ser4", Ser5I9, Ser670, and Thr7", were not phosphorylated. The "lowprobability" PKA sites (those not containing an Arg residue) were not phosphorylated. The results suggest that isolated domains of CFTR developed useful models for investigating the biochemical and structural effects of phosphorylation within CFTR. The mass spectrometry approach in this study should prove useful for defining phosphorylation sites of CFTR in vitro and in vivo.

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Section: E T L H R E S L E G D a P V S W L E T K~f~t G E F G E K R K mentioning

confidence: 99%

“…Only phosphoserine has been detected in CFTR (Cheng et al, 1991). The first nucleotide binding domain (NBDI) contains four Ser residues, Ser4**, Ser4", Ser5", and Se?57 (Fig.…”

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Identification of protein kinase A phosphorylation sites on NBD1 and R domains of CFTR using electrospray mass spectrometry with selective phosphate ion monitoring

et al. 1996

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“…Mutations in the cystic fibrosis transmembrane conductance regulator gene (CFTR) lead to dysfunction or absence of a cAMP regulated chloride channel in the apical membrane of epithelial cells. 1,2 Although various organs of the body are affected, respiratory failure is the cause of death in more than 95% of CF patients. Chronic inflammation, mucus accumulation and persistent bacterial infections cause a progressive decline in lung function.…”

Section: Introductionmentioning

confidence: 99%

“…3 Viral and nonviral vector systems, including cationic liposomes, have been widely used to transfer functional genes into the lungs of laboratory animals. 1,[4][5][6][7][8][9][10][11] In most studies the lung has been directly targeted by administrating the vector via aerosol or intratracheal and intra-nasal injections. Direct intrapulmonary administration can be relatively noninvasive; lung epithelial cells that face the airway lumen are readily accessible and potential problems with ectopic expression and toxicity in organs other than the lung are minimized.…”

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confidence: 99%

Comparison between intratracheal and intravenous administration of liposome–DNA complexes for cystic fibrosis lung gene therapy

Griesenbach

Chonn

Cassady³

et al. 1998

Gene Ther

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Intratracheal (i.t.) and intravenous (i.v.) delivery of DNA-Expression using either mode of administration was maxivector formulations are two strategies to obtain gene transmal 24 h after injection and declined to around 10% of day fer to the lung. It is still uncertain, however, which of these 1 levels 2 weeks after injection. For i.v. delivery of DODAC: two modes of delivery will be more effective in the treat-DOPE-DNA complexes multilamellar vesicles were more ment of cystic fibrosis and other lung diseases. In this effective than large unilamellar vesicles in all organs invesstudy, we attempted to optimize formulations of the cationic tigated. Recombinant DNA could be detected in the distal liposome DODAC:DOPE (dioleoyldimethylammoniumlung region following either route of administration. Howchloride:dioleoylphosphatidylethanolamine) complexed to ever, i.t. administration predominantly led to DNA depoplasmids encoding chloramphenicol acetyltransferase for sition in epithelial cells lining the bronchioles, eg in clara i.t. and i.v. injection into CD-1 mice and compared the two cells, whereas i.v. administration resulted in DNA depomethods. Our results showed that both methods conferred sition in the alveolar region of the lung including type II reporter gene expression in the lung that was significantly alveolar epithelial cells. higher relative to injection of plasmid DNA alone.

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Potentially lethal cystic fibrosis gene variant in the orangutan

Taylor‐Cousar

Evans

Cutting

et al. 2020

American J Primatol

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A syndrome of chronic upper and lower airway disease leading to increased morbidity and mortality occurs primarily in captive orangutans. Similarities in symptoms to the inherited human respiratory disease, cystic fibrosis, led us to hypothesize that orangutan respiratory disease is a result of variants in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene. We identified the nonsense

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Phosphorylation of the R domain by cAMP-dependent protein kinase regulates the CFTR chloride channel

Cited by 141 publications

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Identification of protein kinase A phosphorylation sites on NBD1 and R domains of CFTR using electrospray mass spectrometry with selective phosphate ion monitoring

Identification of protein kinase A phosphorylation sites on NBD1 and R domains of CFTR using electrospray mass spectrometry with selective phosphate ion monitoring

Comparison between intratracheal and intravenous administration of liposome–DNA complexes for cystic fibrosis lung gene therapy

Potentially lethal cystic fibrosis gene variant in the orangutan

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