A5047732079- Cystic Fibrosis Research Advances
- Advanced biosensing and bioanalysis techniques
- Neonatal Respiratory Health Research
- Drug Transport and Resistance Mechanisms
- Ion channel regulation and function
- Inhalation and Respiratory Drug Delivery
- Genomics and Rare Diseases
- DNA and Nucleic Acid Chemistry
- Respiratory viral infections research
- RNA and protein synthesis mechanisms
- Bacterial Genetics and Biotechnology
- Plant Stress Responses and Tolerance
- Legume Nitrogen Fixing Symbiosis
- Ion Channels and Receptors
- Ion Transport and Channel Regulation
- Plant Gene Expression Analysis
- Plant and Biological Electrophysiology Studies
- Cellular transport and secretion
- Lipid Membrane Structure and Behavior
- Trace Elements in Health
- Renal function and acid-base balance
- Plant Reproductive Biology
- Cancer-related gene regulation
- Autism Spectrum Disorder Research
- Infant Nutrition and Health
University College London
2012-2023
Rockefeller University
2002-2018
Semmelweis University
2018
University of London
1997-1999
Wye College
1997-1999
University of Milan
1993-1997
CFTR, the product of gene mutated in cystic fibrosis, is an ATPase that functions as a Cl− channel which bursts openings separate relatively long interburst closed times (τib). Channel gating controlled by phosphorylation and MgATP, but underlying molecular mechanisms remain controversial. To investigate them, we expressed CFTR channels Xenopus oocytes examined, excised patches, how kinetics phosphorylated were affected changes [MgATP], alterations chemical structure activating nucleotide,...
CFTR, the protein defective in cystic fibrosis, functions as a Cl- channel regulated by cAMP-dependent kinase (PKA). CFTR is also an ATPase, comprising two nucleotide-binding domains (NBDs) thought to bind and hydrolyze ATP. In hydrolyzable nucleoside triphosphates, PKA-phosphorylated channels open into bursts, lasting on order of second, from closed (interburst) intervals second or more. To investigate nucleotide interactions underlying gating, we examined photolabeling [alpha32P]8-N3ATP...
CFTR, the ABC protein defective in cystic fibrosis, functions as an anion channel. Once phosphorylated by kinase A, a CFTR channel is opened and closed events at its two cytosolic nucleotide binding domains (NBDs). Formation of head-to-tail NBD1/NBD2 heterodimer, ATP interfacial composite sites between conserved Walker A B motifs one NBD ABC-specific signature sequence other, has been proposed to trigger opening. hydrolysis only catalytically competent site suggested then destabilize dimer...
Cystic fibrosis transmembrane conductance regulator (CFTR) is a chloride channel belonging to the adenosine triphosphate (ATP)-binding cassette (ABC) superfamily. ABC proteins share common molecular mechanism that couples ATP binding and hydrolysis at two nucleotide-binding domains (NBDs) diverse functions. This involves formation of NBD dimers, with bound composite interfacial sites. In CFTR, intramolecular dimerization coupled opening. Channel closing triggered by molecule site 2. Site 1,...
Cystic fibrosis (CF) is a debilitating hereditary disease caused by mutations in the cystic transmembrane conductance regulator (CFTR) gene, which encodes an anion channel. Wild type-CFTR gating non-equilibrium process. After ATP binding, CFTR enters stable open state (O1 ). hydrolysis leads it to short-lived post-hydrolytic (O2 ), from channels close. Here, we use probe mechanism of VX-770, first compound directly targeting protein approved for treatment CF. D1370N and K1250R reduce or...
A central step in the gating of cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel is association its two cytosolic nucleotide-binding domains (NBDs) into a head-to-tail dimer, with nucleotides bound at interface. Channel opening and closing, respectively, are coupled to formation disruption this tight NBD dimer. CFTR an asymmetric adenosine triphosphate (ATP)-binding cassette protein which interfacial-binding sites (composite 1 2) functionally different. During...
Enteroviruses (EVs) are among the most frequent infectious agents in humans worldwide and represent leading cause of upper respiratory tract infections. No drugs for treatment EV infections currently available. Recent studies have also linked infection with pulmonary exacerbations, especially cystic fibrosis (CF) patients, importance this link is probably underestimated. The aim work was to develop a new class multitarget active both as broad-spectrum antivirals correctors F508del-cystic...
Cystic fibrosis (CF) is a debilitating disease caused by mutations in the cystic transmembrane conductance regulator (CFTR) gene, which codes for Cl-/HCO3 - channel. F508del, most common CF-associated mutation, causes both gating and biogenesis defects CFTR protein. This paper describes optimization of two fluorescence assays, capable measuring function cellular localization, their use pilot drug screen.HEK293 cells expressing YFP-F508del-CFTR, halide sensitive YFP tagged to N-terminal CFTR,...
Unique among ABC (ATP-binding cassette) protein family members, CFTR (cystic fibrosis transmembrane conductance regulator), also termed ABCC7, encoded by the gene mutated in cystic patients, functions as an ion channel. Opening and closing of its anion-selective pore are linked to ATP binding hydrolysis at CFTR's two NBDs (nucleotide-binding domains), NBD1 NBD2. Isolated prokaryotic proteins form homodimers upon ATP, but separate after ATP. By combining mutagenesis with single-channel...
The chloride ion channel cystic fibrosis transmembrane conductance regulator (CFTR) displays a typical adenosine trisphosphate (ATP)-binding cassette (ABC) protein architecture comprising two domains, intracellular nucleotide-binding domains (NBDs), and unique regulatory domain. Once phosphorylated in the domain, CFTR channels can open close when supplied with cytosolic ATP. Despite general agreement that formation of head-to-tail NBD dimer drives opening pore, little is known about how ATP...
Gating of the yeast K+ channel encoded by Saccharomyces cerevisiae gene TOK1, unlike other outward-rectifying channels that have been cloned, is promoted membrane voltage (inside positive-going) and repressed extracellular K+. When expressed in Xenopus laevis oocytes, TOK1p current rectified strongly outward, its activation shifting parallel with equilibrium potential when external concentration ([K+]o) was increased above 3 mM. Analysis indicated two kinetic components contributed to...
Key points Cystic fibrosis (CF) is a common genetic disease caused by loss‐of‐function mutations in the cystic transmembrane conductance regulator gene, which encodes channel protein, selective for anions. In lungs, site of most severe symptoms, CF causes abnormal electrolyte transport epithelial cells line airways. Airway ion can be assessed measuring trans‐epithelial potential difference ( V t ) shows characteristic changes individuals. We developed biophysical model human nasal epithelia,...
Cystic fibrosis (CF) is a multiorgan disease caused by mutations of the cystic transmembrane conductance regulator (CFTR). In addition to respiratory impairment due mucus accumulation, viruses and bacteria trigger acute pulmonary exacerbations, accelerating progression mortality rate. Treatment complexity increases with patients' age, simplifying therapeutic regimen represents one key priorities in CF. We have recently reported discovery multitarget compounds able "kill two birds stone"...
Class Ia/b cystic fibrosis transmembrane regulator (CFTR) variants cause severe lung disease in 10% of (CF) patients and are untreatable with small-molecule pharmaceuticals. Genetic replacement CFTR offers a cure, but its effectiveness is limited vivo. We hypothesized that enhancing protein levels (using codon optimization) and/or activity gain-of-function variants) would more effectively restore function to CF bronchial epithelial cells. Three different the were tested: optimized (high...
Deletion of phenylalanine 508 (F508del) in the cystic fibrosis transmembrane conductance regulator (CFTR) anion channel is most common cause fibrosis. The F508 residue located on nucleotide-binding domain 1 (NBD1) contact with cytosolic extensions helices, particular intracellular loop 4 (ICL4). To investigate how absence at this interface impacts CFTR protein, we carried out a mutagenesis scan ICL4 by introducing second-site mutations 11 positions cis F508del. Using an image-based...
Unique among ABC (ATP-binding cassette) protein family members, CFTR (cystic fibrosis transmembrane conductance regulator), also termed ABCC7, encoded by the gene mutated in cystic patients, functions as an ion channel. Opening and closing of its anion-selective pore are linked to ATP binding hydrolysis at CFTR's two NBDs (nucleotide-binding domains), NBD1 NBD2. Isolated prokaryotic proteins form homodimers upon ATP, but separate after ATP. By combining mutagenesis with single-channel...