TY - JOUR AU - Levring, J. AU - Terry, D.S. AU - Kilic, Z. AU - Fitzgerald, G. AU - Blanchard, S. AU - Chen, J. TI - CFTR function, pathology and pharmacology at single-molecule resolution JF - NATURE J2 - NATURE VL - 616 PY - 2023 IS - 7957 SP - 606 EP - 614 PG - 9 SN - 0028-0836 DO - 10.1038/s41586-023-05854-7 UR - https://m2.mtmt.hu/api/publication/34191189 ID - 34191189 N1 - Laboratory of Membrane Biology and Biophysics, The Rockefeller University, New York, NY, United States Department of Structural Biology, St. Jude Children’s Research Hospital, Memphis, TN, United States Department of Physiology and Biophysics, Weill Cornell Medicine, New York, NY, United States Howard Hughes Medical Institute, The Rockefeller University, New York, NY, United States Cited By :1 Export Date: 12 October 2023 CODEN: NATUA Correspondence Address: Chen, J.; Laboratory of Membrane Biology and Biophysics, United States; email: juechen@rockefeller.edu Correspondence Address: Blanchard, S.; Department of Structural Biology, United States; email: Scott.Blanchard@stjude.org Chemicals/CAS: adenosine triphosphatase, 37289-25-1, 9000-83-3; cystic fibrosis transmembrane conductance regulator, 126880-72-6; ivacaftor, 873054-44-5; adenosine triphosphate, 15237-44-2, 56-65-5, 987-65-5; Adenosine Triphosphate; ATP-Binding Cassette Transporters; CFTR protein, human; Cystic Fibrosis Transmembrane Conductance Regulator Funding details: National Institutes of Health, NIH, GM079238 Funding details: Howard Hughes Medical Institute, HHMI Funding details: Rockefeller University Funding text 1: We acknowledge the support from the Single-Molecule Imaging Center at St. Jude Children’s Research Hospital and the Evelyn Gruss Lipper Cryo-EM Resource Center of The Rockefeller University. We thank L. Csanády and I. Chen for their comments on the manuscript. This work was financially supported by HHMI (to J.C.) and the National Institutes of Health (GM079238 to S.B). AB - The cystic fibrosis transmembrane conductance regulator (CFTR) is an anion channel that regulates salt and fluid homeostasis across epithelial membranes1. Alterations in CFTR cause cystic fibrosis, a fatal disease without a cure2,3. Electrophysiological properties of CFTR have been analysed for decades4–6. The structure of CFTR, determined in two globally distinct conformations, underscores its evolutionary relationship with other ATP-binding cassette transporters. However, direct correlations between the essential functions of CFTR and extant structures are lacking at present. Here we combine ensemble functional measurements, single-molecule fluorescence resonance energy transfer, electrophysiology and kinetic simulations to show that the two nucleotide-binding domains (NBDs) of human CFTR dimerize before channel opening. CFTR exhibits an allosteric gating mechanism in which conformational changes within the NBD-dimerized channel, governed by ATP hydrolysis, regulate chloride conductance. The potentiators ivacaftor and GLPG1837 enhance channel activity by increasing pore opening while NBDs are dimerized. Disease-causing substitutions proximal (G551D) or distal (L927P) to the ATPase site both reduce the efficiency of NBD dimerization. These findings collectively enable the framing of a gating mechanism that informs on the search for more efficacious clinical therapies. © 2023, The Author(s). LA - English DB - MTMT ER - TY - JOUR AU - Simon, Márton AU - Csanády, László TI - Optimization of CFTR gating through the evolution of its extracellular loops JF - JOURNAL OF GENERAL PHYSIOLOGY J2 - J GEN PHYSIOL VL - 155 PY - 2023 IS - 4 PG - 16 SN - 0022-1295 DO - 10.1085/jgp.202213264 UR - https://m2.mtmt.hu/api/publication/33636409 ID - 33636409 LA - English DB - MTMT ER - TY - JOUR AU - Simon, Márton AU - Iordanov, Iordan AU - Szöllősi, András AU - Csanády, László TI - Estimating the true stability of the prehydrolytic outward-facing state in an ABC protein. JF - ELIFE J2 - ELIFE VL - 12 PY - 2023 PG - 19 SN - 2050-084X DO - 10.7554/eLife.90736 UR - https://m2.mtmt.hu/api/publication/34232792 ID - 34232792 AB - CFTR, the anion channel mutated in cystic fibrosis patients, is a model ABC protein whose ATP-driven conformational cycle is observable at single-molecule level in patch-clamp recordings. Bursts of CFTR pore openings are coupled to tight dimerization of its two nucleotide-binding domains (NBDs) and in wild-type (WT) channels are mostly terminated by ATP hydrolysis. The slow rate of non-hydrolytic closure - which determines how tightly bursts and ATP hydrolysis are coupled - is unknown, as burst durations of catalytic site mutants span a range of ~200-fold. Here, we show that Walker A mutation K1250A, Walker B mutation D1370N, and catalytic glutamate mutations E1371S and E1371Q all completely disrupt ATP hydrolysis. True non-hydrolytic closing rate of WT CFTR approximates that of K1250A and E1371S. That rate is slowed ~15-fold in E1371Q by a non-native inter-NBD H-bond, and accelerated ~15-fold in D1370N. These findings uncover unique features of the NBD interface in human CFTR. LA - English DB - MTMT ER - TY - JOUR AU - Zeng, Z.W. AU - Linsdell, P. AU - Pomès, R. TI - Molecular dynamics study of Cl− permeation through cystic fibrosis transmembrane conductance regulator (CFTR) JF - CELLULAR AND MOLECULAR LIFE SCIENCES J2 - CELL MOL LIFE SCI VL - 80 PY - 2023 IS - 2 SN - 1420-682X DO - 10.1007/s00018-022-04621-7 UR - https://m2.mtmt.hu/api/publication/33629018 ID - 33629018 N1 - Export Date: 8 February 2023 CODEN: CMLSF Correspondence Address: Pomès, R.; Molecular Medicine, 686 Bay Street, Canada; email: pomes@sickkids.ca Funding details: Compute Canada Funding details: Canadian Institutes of Health Research, IRSC, MOP130461 Funding text 1: We thank Christine Bear for providing feedbacks on early preliminary results. This work was supported by Canadian Institutes of Health Research Grant MOP130461. MD simulations and analyses were enabled by supercomputing resources and support provided by SciNet ( www.scinet.ca ) and Compute Canada ( www.computecanada.ca ). Funding text 2: We thank Christine Bear for providing feedbacks on early preliminary results. This work was supported by Canadian Institutes of Health Research Grant MOP130461. MD simulations and analyses were enabled by supercomputing resources and support provided by SciNet (www.scinet.ca ) and Compute Canada (www.computecanada.ca ). Funding text 3: This work is supported by Canadian Institutes of Health Research grant MOP130461 (R.P.) and SickKids Restracomp (Z.W.Z.). AB - The recent elucidation of atomistic structures of Cl− channel CFTR provides opportunities for understanding the molecular basis of cystic fibrosis. Despite having been activated through phosphorylation and provided with ATP ligands, several near-atomistic cryo-EM structures of CFTR are in a closed state, as inferred from the lack of a continuous passage through a hydrophobic bottleneck region located in the extracellular portion of the pore. Here, we present repeated, microsecond-long molecular dynamics simulations of human CFTR solvated in a lipid bilayer and aqueous NaCl. At equilibrium, Cl− ions enter the channel through a lateral intracellular portal and bind to two distinct cationic sites inside the channel pore but do not traverse the narrow, de-wetted bottleneck. Simulations conducted in the presence of a strong hyperpolarizing electric field led to spontaneous Cl− translocation events through the bottleneck region of the channel, suggesting that the protein relaxed to a functionally open state. Conformational changes of small magnitude involving transmembrane helices 1 and 6 preceded ion permeation through diverging exit routes at the extracellular end of the pore. The pore bottleneck undergoes wetting prior to Cl− translocation, suggesting that it acts as a hydrophobic gate. Although permeating Cl− ions remain mostly hydrated, partial dehydration occurs at the binding sites and in the bottleneck. The observed Cl− pathway is largely consistent with the loci of mutations that alter channel conductance, anion binding, and ion selectivity, supporting the model of the open state of CFTR obtained in the present study. © 2023, The Author(s). LA - English DB - MTMT ER - TY - JOUR AU - Della, Sala A. AU - Prono, G. AU - Hirsch, E. AU - Ghigo, A. TI - Role of Protein Kinase A-Mediated Phosphorylation in CFTR Channel Activity Regulation JF - FRONTIERS IN PHYSIOLOGY J2 - FRONT PHYSIOL VL - 12 PY - 2021 SN - 1664-042X DO - 10.3389/fphys.2021.690247 UR - https://m2.mtmt.hu/api/publication/32111898 ID - 32111898 N1 - Molecular Biotechnology Center, Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy Kither Biotech S.r.l, Turin, Italy Export Date: 24 July 2021 Correspondence Address: Hirsch, E.; Molecular Biotechnology Center, Italy; email: emilio.hirsch@unito.it Correspondence Address: Ghigo, A.; Molecular Biotechnology Center, Italy; email: alessandra.ghigo@unito.it Funding Agency and Grant Number: Italian Cystic Fibrosis Research Foundation (FFC)Ministry of Health, ItalyItalian Cystic Fibrosis Research Foundation [8/2018]; Compagnia di SanpaoloCompagnia di San Paolo [CSTO161109]; Cariplo FoundationFondazione Cariplo [2018-0498]; Roche Foundation (Bando Roche per la Ricerca 2019); Telethon FoundationFondazione Telethon [GGP20079] Funding text: This work was supported by the Italian Cystic Fibrosis Research Foundation (FFC#8/2018 to EH), Compagnia di Sanpaolo (CSTO161109 to EH), Cariplo Foundation (#2018-0498 to EH), Roche Foundation (Bando Roche per la Ricerca 2019 to AG), and Telethon Foundation (GGP20079 to AG). Molecular Biotechnology Center, Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy Kither Biotech S.r.l, Turin, Italy Export Date: 30 August 2021 Correspondence Address: Hirsch, E.; Molecular Biotechnology Center, Italy; email: emilio.hirsch@unito.it Correspondence Address: Ghigo, A.; Molecular Biotechnology Center, Italy; email: alessandra.ghigo@unito.it Molecular Biotechnology Center, Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy Kither Biotech S.r.l, Turin, Italy Export Date: 31 August 2021 Correspondence Address: Hirsch, E.; Molecular Biotechnology Center, Italy; email: emilio.hirsch@unito.it Correspondence Address: Ghigo, A.; Molecular Biotechnology Center, Italy; email: alessandra.ghigo@unito.it Molecular Biotechnology Center, Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy Kither Biotech S.r.l, Turin, Italy Export Date: 9 September 2021 Correspondence Address: Hirsch, E.; Molecular Biotechnology Center, Italy; email: emilio.hirsch@unito.it Correspondence Address: Ghigo, A.; Molecular Biotechnology Center, Italy; email: alessandra.ghigo@unito.it Funding details: CSTO161109 Funding details: Fondazione Telethon, GGP20079 Funding details: Fondazione Cariplo, 2018-0498 Funding details: Fondazione per la Ricerca sulla Fibrosi Cistica, FFC, FFC#8/2018 Funding text 1: This work was supported by the Italian Cystic Fibrosis Research Foundation (FFC#8/2018 to EH), Compagnia di Sanpaolo (CSTO161109 to EH), Cariplo Foundation (#2018-0498 to EH), Roche Foundation (Bando Roche per la Ricerca 2019 to AG), and Telethon Foundation (GGP20079 to AG). Molecular Biotechnology Center, Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy Kither Biotech S.r.l, Turin, Italy Export Date: 13 September 2021 Correspondence Address: Hirsch, E.; Molecular Biotechnology Center, Italy; email: emilio.hirsch@unito.it Correspondence Address: Ghigo, A.; Molecular Biotechnology Center, Italy; email: alessandra.ghigo@unito.it Funding details: CSTO161109 Funding details: Fondazione Telethon, GGP20079 Funding details: Fondazione Cariplo, 2018-0498 Funding details: Fondazione per la Ricerca sulla Fibrosi Cistica, FFC, FFC#8/2018 Funding text 1: This work was supported by the Italian Cystic Fibrosis Research Foundation (FFC#8/2018 to EH), Compagnia di Sanpaolo (CSTO161109 to EH), Cariplo Foundation (#2018-0498 to EH), Roche Foundation (Bando Roche per la Ricerca 2019 to AG), and Telethon Foundation (GGP20079 to AG). Molecular Biotechnology Center, Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy Kither Biotech S.r.l, Turin, Italy Export Date: 14 September 2021 Correspondence Address: Hirsch, E.; Molecular Biotechnology Center, Italy; email: emilio.hirsch@unito.it Correspondence Address: Ghigo, A.; Molecular Biotechnology Center, Italy; email: alessandra.ghigo@unito.it Funding details: CSTO161109 Funding details: Fondazione Telethon, GGP20079 Funding details: Fondazione Cariplo, 2018-0498 Funding details: Fondazione per la Ricerca sulla Fibrosi Cistica, FFC, FFC#8/2018 Funding text 1: This work was supported by the Italian Cystic Fibrosis Research Foundation (FFC#8/2018 to EH), Compagnia di Sanpaolo (CSTO161109 to EH), Cariplo Foundation (#2018-0498 to EH), Roche Foundation (Bando Roche per la Ricerca 2019 to AG), and Telethon Foundation (GGP20079 to AG). LA - English DB - MTMT ER - TY - JOUR AU - Simon, Márton AU - Csanády, László TI - Molecular pathology of the R117H cystic fibrosis mutation is explained by loss of a hydrogen bond JF - ELIFE J2 - ELIFE VL - 10 PY - 2021 PG - 19 SN - 2050-084X DO - 10.7554/eLife.74693 UR - https://m2.mtmt.hu/api/publication/32573094 ID - 32573094 AB - The phosphorylation-activated anion channel cystic fibrosis transmembrane conductance regulator (CFTR) is gated by an ATP hydrolysis cycle at its two cytosolic nucleotide-binding domains, and is essential for epithelial salt-water transport. A large number of CFTR mutations cause cystic fibrosis. Since recent breakthrough in targeted pharmacotherapy, CFTR mutants with impaired gating are candidates for stimulation by potentiator drugs. Thus, understanding the molecular pathology of individual mutations has become important. The relatively common R117H mutation affects an extracellular loop, but nevertheless causes a strong gating defect. Here, we identify a hydrogen bond between the side chain of arginine 117 and the backbone carbonyl group of glutamate 1124 in the cryo-electronmicroscopic structure of phosphorylated, ATP-bound CFTR. We address the functional relevance of that interaction for CFTR gating using macroscopic and microscopic inside-out patch-clamp recordings. Employing thermodynamic double-mutant cycles, we systematically track gating-state-dependent changes in the strength of the R117-E1124 interaction. We find that the H-bond is formed only in the open state, but neither in the short-lived 'flickery' nor in the long-lived 'interburst' closed state. Loss of this H-bond explains the strong gating phenotype of the R117H mutant, including robustly shortened burst durations and strongly reduced intraburst open probability. The findings may help targeted potentiator design. LA - English DB - MTMT ER - TY - JOUR AU - Yeh, Han- I AU - Yu, Ying-Chun AU - Kuo, Pei-Lun AU - Tsai, Chun-Kuang AU - Huang, Hsin-Tuan AU - Hwang, Tzyh-Chang TI - Functional stability of CFTR depends on tight binding of ATP at its degenerate ATP-binding site JF - JOURNAL OF PHYSIOLOGY-LONDON J2 - J PHYSIOL-LONDON VL - 599 PY - 2021 IS - 20 SP - 4625 EP - 4642 PG - 18 SN - 0022-3751 DO - 10.1113/JP281933 UR - https://m2.mtmt.hu/api/publication/32274498 ID - 32274498 N1 - Dalton Cardiovascular Research Center and Department of Medical Pharmacology and Physiology, University of Missouri-Columbia, Columbia, MO 65211, United States Department of Pharmacology, School of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan School of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan Export Date: 18 February 2022 CODEN: JPHYA Correspondence Address: Hwang, T.-C.; Dalton Cardiovascular Research Center and Department of Medical Pharmacology and Physiology, United States; email: hwangt@nycu.edu.tw Chemicals/CAS: adenosine triphosphate, 15237-44-2, 56-65-5, 987-65-5; cystic fibrosis transmembrane conductance regulator, 126880-72-6; Adenosine Triphosphate; CFTR protein, human; Chloride Channels; Cystic Fibrosis Transmembrane Conductance Regulator Funding details: National Institutes of Health, NIH, NIHR01DK55835 Funding details: Cystic Fibrosis Foundation, CFF, Hwang19G0 Funding details: Ministry of Science and Technology, Taiwan, MOST, 109‐2320‐B‐010‐049‐MY2 Funding text 1: This work is supported by the National Institutes of Health (grant NIHR01DK55835), the Cystic Fibrosis Foundation (grant Hwang19G0), and Ministry of Science and Technology, Taiwan (109‐2320‐B‐010‐049‐MY2) to T. ‐C. Hwang. AB - Opening of the cystic fibrosis transmembrane conductance regulator (CFTR) channel is coupled to the motion of its two nucleotide-binding domains: they form a heterodimer sandwiching two functionally distinct ATP-binding sites (sites 1 and 2). While active ATP hydrolysis in site 2 triggers rapid channel closure, the functional role of stable ATP binding in the catalysis-incompetent (or degenerate) site 1, a feature conserved in many other ATP-binding cassette (ABC) transporter proteins, remains elusive. Here, we found that CFTR loses its prompt responsiveness to ATP after the channel is devoid of ATP for tens to hundreds of seconds. Mutants with weakened ATP binding in site 1 and the most prevalent disease-causing mutation, F508del, are more vulnerable to ATP depletion. In contrast, strengthening ligand binding in site 1 with N-6-(2-phenylethyl)-ATP, a high-affinity ATP analogue, or abolishing ATP hydrolysis in site 2 by the mutation D1370N, helps sustain a durable function of the otherwise unstable mutant channels. Thus, tight binding of ATP in the degenerate ATP-binding site is crucial to the functional stability of CFTR. Small molecules targeting site 1 may bear therapeutic potential to overcome the membrane instability of F508del-CFTR. LA - English DB - MTMT ER - TY - JOUR AU - Stockner, T. AU - Gradisch, R. AU - Schmitt, L. TI - The role of the degenerate nucleotide binding site in type I ABC exporters JF - FEBS LETTERS J2 - FEBS LETT VL - 594 PY - 2020 IS - 23 SP - 3815 EP - 3838 PG - 24 SN - 0014-5793 DO - 10.1002/1873-3468.13997 UR - https://m2.mtmt.hu/api/publication/31732979 ID - 31732979 N1 - Institute of Pharmacology, Center for Physiology and Pharmacology, Medical University of Vienna, Vienna, Austria Institute of Biochemistry, Heinrich Heine University, Düsseldorf, Germany Cited By :3 Export Date: 14 September 2021 CODEN: FEBLA Correspondence Address: Stockner, T.; Institute of Pharmacology, Austria; email: thomas.stockner@meduniwien.ac.at Correspondence Address: Schmitt, L.; Institute of Biochemistry, Germany; email: lutz.schmitt@hhu.de Chemicals/CAS: ABC transporter subfamily B, 149200-37-3, 208997-77-7; cystic fibrosis transmembrane conductance regulator, 126880-72-6; multidrug resistance associated protein 1; adenosine triphosphate, 15237-44-2, 56-65-5, 987-65-5; Adenosine Triphosphate; ATP-Binding Cassette Transporters Funding details: Deutsche Forschungsgemeinschaft, DFG, CRC 1208, Schm1279/13‐1, Schm1279/13‐1 CRC 1208 Funding details: Austrian Science Fund, FWF, 32017, FWF‐SFB035‐24 Funding text 1: This work was supported by grants from the Austrian Science Fund (FWF‐SFB035‐24 and Stand‐Alone Project 32017) to TS and DFG (CRC 1208, project A01, and Schm1279/13‐1 CRC 1208, project A01, and Schm1279/13‐1) to LS. Institute of Pharmacology, Center for Physiology and Pharmacology, Medical University of Vienna, Vienna, Austria Institute of Biochemistry, Heinrich Heine University, Düsseldorf, Germany Cited By :3 Export Date: 15 September 2021 CODEN: FEBLA Correspondence Address: Stockner, T.; Institute of Pharmacology, Austria; email: thomas.stockner@meduniwien.ac.at Correspondence Address: Schmitt, L.; Institute of Biochemistry, Germany; email: lutz.schmitt@hhu.de Chemicals/CAS: ABC transporter subfamily B, 149200-37-3, 208997-77-7; cystic fibrosis transmembrane conductance regulator, 126880-72-6; multidrug resistance associated protein 1; adenosine triphosphate, 15237-44-2, 56-65-5, 987-65-5; Adenosine Triphosphate; ATP-Binding Cassette Transporters Funding details: Deutsche Forschungsgemeinschaft, DFG, CRC 1208, Schm1279/13‐1, Schm1279/13‐1 CRC 1208 Funding details: Austrian Science Fund, FWF, 32017, FWF‐SFB035‐24 Funding text 1: This work was supported by grants from the Austrian Science Fund (FWF‐SFB035‐24 and Stand‐Alone Project 32017) to TS and DFG (CRC 1208, project A01, and Schm1279/13‐1 CRC 1208, project A01, and Schm1279/13‐1) to LS. AB - ATP-binding cassette (ABC) transporters are fascinating molecular machines that are capable of transporting a large variety of chemically diverse compounds. The energy required for translocation is derived from binding and hydrolysis of ATP. All ABC transporters share a basic architecture and are composed of two transmembrane domains and two nucleotide binding domains (NBDs). The latter harbor all conserved sequence motifs that hallmark the ABC transporter superfamily. The NBDs form the nucleotide binding sites (NBSs) in their interface. Transporters with two active NBSs are called canonical transporters, while ABC exporters from eukaryotic organisms, including humans, frequently have a degenerate NBS1 containing noncanonical residues that strongly impair ATP hydrolysis. Here, we summarize current knowledge on degenerate ABC transporters. By integrating structural information with biophysical and biochemical evidence of asymmetric function, we develop a model for the transport cycle of degenerate ABC transporters. We will elaborate on the unclear functional advantages of a degenerate NBS. © 2020 The Authors. FEBS Letters published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies LA - English DB - MTMT ER - TY - JOUR AU - Chin, Stephanie AU - Ramjeesingh, Mohabir AU - Hung, Maurita AU - Ereno-Oreba, June AU - Cui, Hong AU - Laselva, Onofrio AU - Julien, Jean-Philippe AU - Bear, Christine E. TI - Cholesterol Interaction Directly Enhances Intrinsic Activity of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) JF - CELLS J2 - CELLS-BASEL VL - 8 PY - 2019 IS - 8 PG - 16 SN - 2073-4409 DO - 10.3390/cells8080804 UR - https://m2.mtmt.hu/api/publication/30881158 ID - 30881158 N1 - Export Date: 3 August 2022 AB - The recent cryo-electron microscopy structures of zebrafish and the human cystic fibrosis transmembrane conductance regulator (CFTR) provided unprecedented insights into putative mechanisms underlying gating of its anion channel activity. Interestingly, despite predictions based on channel activity measurements in biological membranes, the structure of the detergent purified, phosphorylated, and ATP-bound human CFTR protein did not reveal a stably open conduction pathway. This study tested the hypothesis that the functional properties of the detergent solubilized CFTR protein used for structural determinations are different from those exhibited by CFTR purified under conditions that retain associated lipids native to the membrane. It was found that CFTR purified together with phospholipids and cholesterol using amphipol: A8-35, exhibited higher rates of catalytic activity, phosphorylation dependent channel activation and potentiation by the therapeutic compound, ivacaftor, than did CFTR purified in detergent. The catalytic activity of phosphorylated CFTR detergent micelles was rescued by the addition of phospholipids plus cholesterol, but not by phospholipids alone, arguing for a specific role for cholesterol in modulating this function. In summary, these studies highlight the importance of lipid interactions in the intrinsic activities and pharmacological potentiation of CFTR. LA - English DB - MTMT ER - TY - JOUR AU - Csanády, László AU - Vergani, Paola AU - Gadsby, David C TI - STRUCTURE, GATING, AND REGULATION OF THE CFTR ANION CHANNEL. JF - PHYSIOLOGICAL REVIEWS J2 - PHYSIOL REV VL - 99 PY - 2019 IS - 1 SP - 707 EP - 738 PG - 32 SN - 0031-9333 DO - 10.1152/physrev.00007.2018 UR - https://m2.mtmt.hu/api/publication/30387986 ID - 30387986 N1 - Department of Medical Biochemistry, Semmelweis UniversityBudapest, Hungary MTA-SE Ion Channel Research GroupBudapest, Hungary Department of Neuroscience, Physiology and Pharmacology, University College London, London, United Kingdom Laboratory of Cardiac/Membrane Physiology, The Rockefeller University, New York, NY, United States Cited By :24 Export Date: 2 July 2020 CODEN: PHREA Chemicals/CAS: adenosine triphosphate, 15237-44-2, 56-65-5, 987-65-5; cyclic AMP dependent protein kinase; cystic fibrosis transmembrane conductance regulator, 126880-72-6; Adenosine Triphosphate; Anions; Cystic Fibrosis Transmembrane Conductance Regulator Funding details: Cystic Fibrosis Trust, CF Funding text 1: Supported by Cystic Fibrosis Trust Project no. SRC 005 and Sparks Grant reference no. 15UCL04 (to P. Vergani), and Hungarian Academy of Sciences Lendület Grant LP2017–14/2017 and Cystic Fibrosis Foundation Research Grant CSANAD17G0 (to L. Csanády). Department of Medical Biochemistry, Semmelweis UniversityBudapest, Hungary MTA-SE Ion Channel Research GroupBudapest, Hungary Department of Neuroscience, Physiology and Pharmacology, University College London, London, United Kingdom Laboratory of Cardiac/Membrane Physiology, The Rockefeller University, New York, NY, United States Cited By :60 Export Date: 11 August 2021 CODEN: PHREA Funding Agency and Grant Number: Cystic Fibrosis Trust [SRC 005]; Sparks Grant [15UCL04]; Hungarian Academy of Sciences Lendulet Grant [LP2017-14/2017]; Cystic Fibrosis Foundation Research Grant [CSANAD17G0] Funding text: Supported by Cystic Fibrosis Trust Project no. SRC 005 and Sparks Grant reference no. 15UCL04 (to P. Vergani), and Hungarian Academy of Sciences Lendulet Grant LP2017-14/2017 and Cystic Fibrosis Foundation Research Grant CSANAD17G0 (to L. Csanady). Department of Medical Biochemistry, Semmelweis UniversityBudapest, Hungary MTA-SE Ion Channel Research GroupBudapest, Hungary Department of Neuroscience, Physiology and Pharmacology, University College London, London, United Kingdom Laboratory of Cardiac/Membrane Physiology, The Rockefeller University, New York, NY, United States Cited By :60 Export Date: 30 August 2021 CODEN: PHREA Department of Medical Biochemistry, Semmelweis UniversityBudapest, Hungary MTA-SE Ion Channel Research GroupBudapest, Hungary Department of Neuroscience, Physiology and Pharmacology, University College London, London, United Kingdom Laboratory of Cardiac/Membrane Physiology, The Rockefeller University, New York, NY, United States Cited By :60 Export Date: 31 August 2021 CODEN: PHREA AB - The cystic fibrosis transmembrane conductance regulator (CFTR) belongs to the ATP binding cassette (ABC) transporter superfamily but functions as an anion channel crucial for salt and water transport across epithelial cells. CFTR dysfunction, because of mutations, causes cystic fibrosis (CF). The anion-selective pore of the CFTR protein is formed by its two transmembrane domains (TMDs) and regulated by its cytosolic domains: two nucleotide binding domains (NBDs) and a regulatory (R) domain. Channel activation requires phosphorylation of the R domain by cAMP-dependent protein kinase (PKA), and pore opening and closing (gating) of phosphorylated channels is driven by ATP binding and hydrolysis at the NBDs. This review summarizes available information on structure and mechanism of the CFTR protein, with a particular focus on atomic-level insight gained from recent cryo-electron microscopic structures and on the molecular mechanisms of channel gating and its regulation. The pharmacological mechanisms of small molecules targeting CFTR's ion channel function, aimed at treating patients suffering from CF and other diseases, are briefly discussed. LA - English DB - MTMT ER - TY - JOUR AU - Rey, Michael M. AU - Bonk, Michael P. AU - Hadjiliadis, Denis TI - Cystic Fibrosis: Emerging Understanding and Therapies JF - ANNUAL REVIEW OF MEDICINE J2 - ANNU REV MED VL - 70 PY - 2019 SP - 197 EP - 210 PG - 14 SN - 0066-4219 DO - 10.1146/annurev-med-112717-094536 UR - https://m2.mtmt.hu/api/publication/30658945 ID - 30658945 N1 - Cited By :4 Export Date: 13 February 2020 CODEN: ARMCA Correspondence Address: Hadjiliadis, D.; Division of Pulmonary, Allergy and Critical Care, Perelman School of Medicine, University of PennsylvaniaUnited States; email: Denis.Hadjiliadis@uphs.upenn.edu Chemicals/CAS: cystic fibrosis transmembrane conductance regulator, 126880-72-6; ivacaftor, 873054-44-5; lumacaftor, 936727-05-8; Aminophenols; Aminopyridines; Benzodioxoles; CFTR protein, human; Cystic Fibrosis Transmembrane Conductance Regulator; ivacaftor; lumacaftor; Quinolones Funding details: Novartis Funding details: Cystic Fibrosis Foundation Funding text 1: M.R.R. and M.P.B. are not aware of any affiliations, memberships, funding, or financial holdings that might be perceived as affecting the objectivity of this review. D.H. has received funding from the Cystic Fibrosis Foundation for conducting studies and for the annual center grant and has participated in CF Foundation Guideline development; he has also received a grant from Novartis for conduct of a multicenter clinical trial. AB - Cystic fibrosis (CF) is the most common life-limiting genetic disease in Caucasian patients. Continued advances have led to improved survival, and adults with CF now outnumber children. As our understanding of the disease improves, new therapies have emerged that improve the basic defect, enabling patient-specific treatment and improved outcomes. However, recurrent ex-acerbations continue to lead to morbidity and mortality, and new pathogens have been identified that may lead to worse outcomes. In addition, new complications, such as CF-related diabetes and increased risk of gastrointestinal cancers, are creating new challenges in management. For patients with end-stage disease, lung transplantation has remained one of the few treatment options, but challenges in identifying the most appropriate patients remain. LA - English DB - MTMT ER - TY - JOUR AU - Martin, SL AU - Saint-Criq, V AU - Hwang, TC AU - Csanády, László TI - Ion channels as targets to treat cystic fibrosis lung disease JF - JOURNAL OF CYSTIC FIBROSIS J2 - J CYST FIBROS VL - 17 PY - 2018 IS - 2 SP - S22 EP - S27 PG - 6 SN - 1569-1993 DO - 10.1016/j.jcf.2017.10.006 UR - https://m2.mtmt.hu/api/publication/3287704 ID - 3287704 N1 - Cited By :4 Export Date: 9 September 2019 CODEN: JCFOA Correspondence Address: Martin, S.L.; School of Pharmacy, Queen's University Belfast, 97 Lisburn Road, United Kingdom; email: l.martin@qub.ac.uk Chemicals/CAS: cystic fibrosis transmembrane conductance regulator, 126880-72-6; Cystic Fibrosis Transmembrane Conductance Regulator Funding details: Transport Funding details: National Institutes of Health, R01DK55835 Funding details: Cystic Fibrosis Foundation, Hwang11P0, CSANAD17G0 Funding details: PJ552, PJ559 Funding details: Medical Research Council, INOVCF/SRC003, MC_PC_141113 Funding details: Invest Northern Ireland, POC600 Funding text 1: We acknowledge the support of the organising committee of the European Cystic Fibrosis Basic Science conference 2017 and the contributions that were also made by Drs Paolo Scudieri (Telethon Institute of Genetics and Medicine) and Génesis Vega (Centro de Estudios Cientificos, Valdivia, Chile) to Symposium 6 (Cell Physiology and Ion Transport) of the conference. SLM's work has received financial support from the CF Trust, UK (PJ552 and PJ559), Invest Northern Ireland (POC600) and the Medical Research Council (MC_PC_141113); VSC is currently funded by a Strategic Research Centre grant (INOVCF/SRC003) from the CF Trust UK; TCH by research grants from the NIH (R01DK55835) and Cystic Fibrosis Foundation (Hwang11P0) and LC by a Research Grant from the Cystic Fibrosis Foundation (CSANAD17G0). AB - Lung health relies on effective mucociliary clearance and innate immune defence mechanisms. In cystic fibrosis (CF), an imbalance in ion transport due to an absence of chloride ion secretion, caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) and a concomitant sodium hyperabsorption, caused by dyregulation of the epithelial sodium channel (ENaC), results in mucus stasis which predisposes the lungs to cycles of chronic infection and inflammation leading to lung function decline. An increased understanding of CFTR structure and function has provided opportunity for the development of a number of novel modulators targeting mutant CFTR however, it is important to also consider other ion channels and transporters present in the airways as putative targets for drug development. In this review, we discuss recent advances in CFTR biology which will contribute to further drug discovery in the field. We also examine developments to inhibit the epithelial sodium channel (ENaC) and potentially activate alternative chloride channels and transporters as a multi-tracked strategy to hydrate CF airways and restore normal mucociliary clearance mechanisms in a manner independent of CFTR mutation. LA - English DB - MTMT ER - TY - JOUR AU - Trowitzsch, S. AU - Tampé, R. TI - ABC Transporters in Dynamic Macromolecular Assemblies JF - JOURNAL OF MOLECULAR BIOLOGY J2 - J MOL BIOL VL - 430 PY - 2018 IS - 22 SP - 4481 EP - 4495 PG - 15 SN - 0022-2836 DO - 10.1016/j.jmb.2018.07.028 UR - https://m2.mtmt.hu/api/publication/30363065 ID - 30363065 N1 - Cited By :1 Export Date: 27 December 2018 CODEN: JMOBA Correspondence Address: Trowitzsch, S.; Institute of Biochemistry, Biocenter, Goethe University Frankfurt, Max-von-Laue Str. 9, Germany; email: trowitzsch@biochem.uni-frankfurt.de Chemicals/CAS: adenosine triphosphate, 15237-44-2, 56-65-5, 987-65-5 Funding details: Deutsche Forschungsgemeinschaft, DFG, SFB 807 Funding details: Deutsche Forschungsgemeinschaft, DFG, GRK 1986 Funding text 1: This research was supported by the German Research Foundation ( SFB 807 and GRK 1986 to R.T.). We are grateful to all laboratory members for helpful comments on the manuscript. Although we have tried to cover the many different aspects of ABC transporters in this contribution, particularly in regard to the PLC, we apologize to any colleagues whose work might not have been adequately cited or extensively discussed. Cited By :20 Export Date: 9 September 2021 CODEN: JMOBA Correspondence Address: Trowitzsch, S.; Institute of Biochemistry, Max-von-Laue Str. 9, Germany; email: trowitzsch@biochem.uni-frankfurt.de Chemicals/CAS: adenosine triphosphate, 15237-44-2, 56-65-5, 987-65-5; ATP-Binding Cassette Transporters; Multiprotein Complexes Funding details: Deutsche Forschungsgemeinschaft, DFG, GRK 1986, SFB 807 Funding text 1: This research was supported by the German Research Foundation ( SFB 807 and GRK 1986 to R.T.). We are grateful to all laboratory members for helpful comments on the manuscript. Although we have tried to cover the many different aspects of ABC transporters in this contribution, particularly in regard to the PLC, we apologize to any colleagues whose work might not have been adequately cited or extensively discussed. LA - English DB - MTMT ER - TY - JOUR AU - Callebaut, Isabelle AU - Hoffmann, Brice AU - Lehn, Pierre AU - Mornon, Jean-Paul TI - Molecular modelling and molecular dynamics of CFTR JF - CELLULAR AND MOLECULAR LIFE SCIENCES J2 - CELL MOL LIFE SCI VL - 74 PY - 2017 IS - 1 SP - 3 EP - 22 PG - 20 SN - 1420-682X DO - 10.1007/s00018-016-2385-9 UR - https://m2.mtmt.hu/api/publication/26407981 ID - 26407981 N1 - UMR CNRS 7590, Museum National d’Histoire Naturelle, IRD UMR 206, IUC, Case 115, IMPMC, Sorbonne Universités, UPMC Univ Paris 06, 4 Place Jussieu, Paris Cedex 05, 75005, France INSERM U1078, SFR ScInBioS, Université de Bretagne Occidentale, Brest, France Cited By :24 Export Date: 10 August 2021 CODEN: CMLSF Correspondence Address: Callebaut, I.; UMR CNRS 7590, 4 Place Jussieu, France; email: isabelle.callebaut@impmc.upmc.fr UMR CNRS 7590, Museum National d’Histoire Naturelle, IRD UMR 206, IUC, Case 115, IMPMC, Sorbonne Universités, UPMC Univ Paris 06, 4 Place Jussieu, Paris Cedex 05, 75005, France INSERM U1078, SFR ScInBioS, Université de Bretagne Occidentale, Brest, France Cited By :24 Export Date: 11 August 2021 CODEN: CMLSF Correspondence Address: Callebaut, I.; UMR CNRS 7590, 4 Place Jussieu, France; email: isabelle.callebaut@impmc.upmc.fr Funding Agency and Grant Number: Association "Vaincre La Mucoviscidose'' (Paris, France); GENCI [2014-077206, 2015-077206, 2016-077206] Funding text: We thank the Association "Vaincre La Mucoviscidose'' (Paris, France) for constant support. This work was granted access to the HPC resources of IDRIS/CINES under the allocations 2014-077206, 2015-077206 and 2016-077206 made by GENCI. UMR CNRS 7590, Museum National d’Histoire Naturelle, IRD UMR 206, IUC, Case 115, IMPMC, Sorbonne Universités, UPMC Univ Paris 06, 4 Place Jussieu, Paris Cedex 05, 75005, France INSERM U1078, SFR ScInBioS, Université de Bretagne Occidentale, Brest, France Cited By :24 Export Date: 27 August 2021 CODEN: CMLSF Correspondence Address: Callebaut, I.; UMR CNRS 7590, 4 Place Jussieu, France; email: isabelle.callebaut@impmc.upmc.fr UMR CNRS 7590, Museum National d’Histoire Naturelle, IRD UMR 206, IUC, Case 115, IMPMC, Sorbonne Universités, UPMC Univ Paris 06, 4 Place Jussieu, Paris Cedex 05, 75005, France INSERM U1078, SFR ScInBioS, Université de Bretagne Occidentale, Brest, France Cited By :24 Export Date: 31 August 2021 CODEN: CMLSF Correspondence Address: Callebaut, I.; UMR CNRS 7590, 4 Place Jussieu, France; email: isabelle.callebaut@impmc.upmc.fr LA - English DB - MTMT ER - TY - JOUR AU - Chin, Stephanie AU - Yang, Donghe AU - Miles, Andrew J AU - Eckford, Paul D W AU - Molinski, Steven AU - Wallace, B A AU - Bear, Christine E TI - Attenuation of Phosphorylation-dependent Activation of Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) by Disease-causing Mutations at the Transmission Interface JF - JOURNAL OF BIOLOGICAL CHEMISTRY J2 - J BIOL CHEM VL - 292 PY - 2017 IS - 5 SP - 1988 EP - 1999 PG - 12 SN - 0021-9258 DO - 10.1074/jbc.M116.762633 UR - https://m2.mtmt.hu/api/publication/26573010 ID - 26573010 N1 - Megjegyzés-26666966 N1 Funding details: CIHR, Canadian Institutes of Health Research N1 Funding text: This work was supported by grants from NSERC (Natural Science and Engineering Research Council; to S. C.) and CIHR (Canadian Institutes of Health Research). This work was also supported by a grant from the UK Biotechnology and Biological Sciences Research Council (to B. A. W.) and by beamtime grants from the ISA synchrotron (to B. A. W.). The authors declare that they have no conflicts of interest with the contents of this article. Peter Gilgan Centre for Research and Learning, Programme of Molecular Structure and Function, Hospital for Sick Children, 686 Bay St., Rm. 209420 U-West, Toronto, ON M5G 0A4, Canada Department of Biochemistry, University of Toronto, Toronto, Canada Institute of Structural and Molecular Biology, Birkbeck College, University of London, London, WC1E 7HX, United Kingdom Department of Physiology, University of Toronto, Toronto, Canada Cited By :3 Export Date: 27 December 2018 CODEN: JBCHA Correspondence Address: Bear, C.E.; Peter Gilgan Centre for Research and Learning, Programme of Molecular Structure and Function, Hospital for Sick Children, 686 Bay St., Rm. 209420 U-West, Canada; email: bear@sickkids.ca Chemicals/CAS: cyclic AMP dependent protein kinase; cystic fibrosis transmembrane conductance regulator, 126880-72-6; iodide, 20461-54-5; tryptophan, 6912-86-3, 73-22-3; CFTR protein, human; Cyclic AMP-Dependent Protein Kinases; Cystic Fibrosis Transmembrane Conductance Regulator Funding details: Natural Sciences and Engineering Research Council of Canada, NSERC Funding details: Biotechnology and Biological Sciences Research Council, BBSRC Funding details: Canadian Institutes of Health Research, CIHR Funding details: Canadian Institutes of Health Research, CIHR Funding details: Natural Sciences and Engineering Research Council of Canada, NSERC Funding details: Iowa Soybean Association, ISA Funding text 1: This work was supported by grants from NSERC (Natural Science and Engineering Research Council; to S. C.) and CIHR (Canadian Institutes of Health Research). This work was also supported by a grant from the UK Biotechnology and Biological Sciences Research Council (to B. A. W.) and by beamtime grants from the ISA synchrotron (to B. A. W.). The authors declare that they have no conflicts of interest with the contents of this article. Peter Gilgan Centre for Research and Learning, Programme of Molecular Structure and Function, Hospital for Sick Children, 686 Bay St., Rm. 209420 U-West, Toronto, ON M5G 0A4, Canada Department of Biochemistry, University of Toronto, Toronto, Canada Institute of Structural and Molecular Biology, Birkbeck College, University of London, London, WC1E 7HX, United Kingdom Department of Physiology, University of Toronto, Toronto, Canada Cited By :6 Export Date: 10 August 2021 CODEN: JBCHA Correspondence Address: Bear, C.E.; Peter Gilgan Centre for Research and Learning, 686 Bay St., Rm. 209420 U-West, Canada; email: bear@sickkids.ca Funding Agency and Grant Number: NSERC (Natural Science and Engineering Research Council)Natural Sciences and Engineering Research Council of Canada (NSERC); CIHR (Canadian Institutes of Health Research)Canadian Institutes of Health Research (CIHR); UK Biotechnology and Biological Sciences Research CouncilUK Research & Innovation (UKRI)Biotechnology and Biological Sciences Research Council (BBSRC); ISA synchrotron; Biotechnology and Biological Sciences Research CouncilUK Research & Innovation (UKRI)Biotechnology and Biological Sciences Research Council (BBSRC) [BBS/B/02959, BB/J019135/1] Funding Source: researchfish Funding text: This work was supported by grants from NSERC (Natural Science and Engineering Research Council; to S.C.) and CIHR (Canadian Institutes of Health Research). This work was also supported by a grant from the UK Biotechnology and Biological Sciences Research Council (to B.A.W.) and by beamtime grants from the ISA synchrotron (to B.A.W.). The authors declare that they have no conflicts of interest with the contents of this article. Peter Gilgan Centre for Research and Learning, Programme of Molecular Structure and Function, Hospital for Sick Children, 686 Bay St., Rm. 209420 U-West, Toronto, ON M5G 0A4, Canada Department of Biochemistry, University of Toronto, Toronto, Canada Institute of Structural and Molecular Biology, Birkbeck College, University of London, London, WC1E 7HX, United Kingdom Department of Physiology, University of Toronto, Toronto, Canada Cited By :6 Export Date: 31 August 2021 CODEN: JBCHA Correspondence Address: Bear, C.E.; Peter Gilgan Centre for Research and Learning, 686 Bay St., Rm. 209420 U-West, Canada; email: bear@sickkids.ca AB - Cystic fibrosis transmembrane conductance regulator (CFTR) is a multidomain membrane protein that functions as a phosphorylation-regulated anion channel. The interface between its two cytosolic nucleotide binding domains and coupling helices conferred by intracellular loops extending from the channel pore domains has been referred to as a transmission interface and is thought to be critical for the regulated channel activity of CFTR. Phosphorylation of the regulatory domain of CFTR by protein kinase A (PKA) is required for its channel activity. However, it was unclear if phosphorylation modifies the transmission interface. Here, we studied purified full-length CFTR protein using spectroscopic techniques to determine the consequences of PKA-mediated phosphorylation. Synchrotron radiation circular dichroism spectroscopy confirmed that purified full-length wild-type CFTR is folded and structurally responsive to phosphorylation. Intrinsic tryptophan fluorescence studies of CFTR showed that phosphorylation reduced iodide-mediated quenching, consistent with an effect of phosphorylation in burying tryptophans at the transmission interface. Importantly, the rate of phosphorylation-dependent channel activation was compromised by the introduction of disease-causing mutations in either of the two coupling helices predicted to interact with nucleotide binding domain 1 at the interface. Together, these results suggest that phosphorylation modifies the interface between the catalytic and pore domains of CFTR and that this modification facilitates CFTR channel activation. LA - English DB - MTMT ER - TY - JOUR AU - Chin, Stephanie AU - Hung, Maurita AU - Bear, Christine E TI - Current insights into the role of PKA phosphorylation in CFTR channel activity and the pharmacological rescue of cystic fibrosis disease-causing mutants JF - CELLULAR AND MOLECULAR LIFE SCIENCES J2 - CELL MOL LIFE SCI VL - 74 PY - 2017 IS - 1 SP - 57 EP - 66 PG - 10 SN - 1420-682X DO - 10.1007/s00018-016-2388-6 UR - https://m2.mtmt.hu/api/publication/26409492 ID - 26409492 N1 - Cited By :9 Export Date: 10 August 2021 CODEN: CMLSF Correspondence Address: Bear, C.E.; Programme of Molecular Structure and Function, Canada; email: bear@sickkids.ca Funding Agency and Grant Number: NSERC scholarship (PGS-D); Canadian Institutes of Health ResearchCanadian Institutes of Health Research (CIHR); Cystic Fibrosis Canada Funding text: S. Chin was supported by a NSERC scholarship (PGS-D) and the research activities in the Bear Lab supported in part by the Canadian Institutes of Health Research and Cystic Fibrosis Canada. Cited By :9 Export Date: 27 August 2021 CODEN: CMLSF Correspondence Address: Bear, C.E.; Programme of Molecular Structure and Function, Canada; email: bear@sickkids.ca Cited By :9 Export Date: 30 August 2021 CODEN: CMLSF Correspondence Address: Bear, C.E.; Programme of Molecular Structure and Function, Canada; email: bear@sickkids.ca Cited By :9 Export Date: 31 August 2021 CODEN: CMLSF Correspondence Address: Bear, C.E.; Programme of Molecular Structure and Function, Canada; email: bear@sickkids.ca LA - English DB - MTMT ER - TY - JOUR AU - Kuehn, Frank AU - Kuehn, Cornelia AU - Lueckhoff, Andreas TI - Different Principles of ADP-Ribose-Mediated Activation and Opposite Roles of the NUDT9 Homology Domain in the TRPM2 Orthologs of Man and Sea Anemone JF - FRONTIERS IN PHYSIOLOGY J2 - FRONT PHYSIOL VL - 8 PY - 2017 PG - 14 SN - 1664-042X DO - 10.3389/fphys.2017.00879 UR - https://m2.mtmt.hu/api/publication/27095255 ID - 27095255 LA - English DB - MTMT ER - TY - JOUR AU - Liu, F AU - Zhang, Z AU - Csanády, László AU - Gadsby, DC AU - Chen, J TI - Molecular Structure of the Human CFTR Ion Channel JF - CELL J2 - CELL VL - 169 PY - 2017 IS - 1 SP - 85 EP - 95.e8 SN - 0092-8674 DO - 10.1016/j.cell.2017.02.024 UR - https://m2.mtmt.hu/api/publication/3210856 ID - 3210856 N1 - Cited By :244 Export Date: 9 March 2022 CODEN: CELLB Correspondence Address: Chen, J.; Laboratory of Membrane Biophysics and Biology, 1230 York Avenue, United States Chemicals/CAS: adenosine triphosphatase, 37289-25-1, 9000-83-3; adenosine triphosphate, 15237-44-2, 56-65-5, 987-65-5; arginine, 1119-34-2, 15595-35-4, 7004-12-8, 74-79-3; cyclic AMP dependent protein kinase; cysteine, 4371-52-2, 52-89-1, 52-90-4; cystic fibrosis transmembrane conductance regulator, 126880-72-6; lysine, 56-87-1, 6899-06-5, 70-54-2; multidrug resistance associated protein 1; proline, 147-85-3, 7005-20-1; protein, 67254-75-5; serine, 56-45-1, 6898-95-9; Adenosine Triphosphate; CFTR protein, human; CFTR protein, zebrafish; Cystic Fibrosis Transmembrane Conductance Regulator; Zebrafish Proteins Funding details: LP2012-39/2012 Funding details: Howard Hughes Medical Institute, HHMI Funding details: Cystic Fibrosis Foundation, CFF, CSANAD15G0 Funding details: Rockefeller University Funding text 1: We thank Eric Gouaux for the expression vector, Mark Ebrahim and Johanna Sotiris at the Rockefeller Evelyn Gruss Lipper Cryo-Electron Microscopy Resource Center for assistance in data collection, and Sarah McCarry for editing this manuscript. This work is supported by the Rockefeller University (to J.C. and D.C.G), the Howard Hughes Medical Institute (to J.C.), and MTA-Momentum (LP2012-39/2012) and Cystic Fibrosis Foundation (CSANAD15G0) grants (to L.C.). AB - The cystic fibrosis transmembrane conductance regulator (CFTR) is an ATP-binding cassette (ABC) transporter that uniquely functions as an ion channel. Here, we present a 3.9 Å structure of dephosphorylated human CFTR without nucleotides, determined by electron cryomicroscopy (cryo-EM). Close resemblance of this human CFTR structure to zebrafish CFTR under identical conditions reinforces its relevance for understanding CFTR function. The human CFTR structure reveals a previously unresolved helix belonging to the R domain docked inside the intracellular vestibule, precluding channel opening. By analyzing the sigmoid time course of CFTR current activation, we propose that PKA phosphorylation of the R domain is enabled by its infrequent spontaneous disengagement, which also explains residual ATPase and gating activity of dephosphorylated CFTR. From comparison with MRP1, a feature distinguishing CFTR from all other ABC transporters is the helix-loop transition in transmembrane helix 8, which likely forms the structural basis for CFTR's channel function. © 2017 Elsevier Inc. LA - English DB - MTMT ER - TY - JOUR AU - Papadaki, Georgia F AU - Amillis, Sotiris AU - Diallinas, George TI - Substrate Specificity of the FurE Transporter Is Determined by Cytoplasmic Terminal Domain Interactions JF - GENETICS J2 - GENETICS VL - 207 PY - 2017 IS - 4 SP - 1387 EP - 1400 PG - 14 SN - 0016-6731 DO - 10.1534/genetics.117.300327 UR - https://m2.mtmt.hu/api/publication/27095230 ID - 27095230 N1 - Cited By :1 Export Date: 27 December 2018 CODEN: GENTA Correspondence Address: Diallinas, G.; Department of Biology, National and Kapodistrian University of Athens, Panepistimioupolis, Greece; email: diallina@biol.uoa.gr Chemicals/CAS: allantoin, 97-59-6; ammonia, 14798-03-9, 51847-23-5, 7664-41-7; uracil, 66-22-8; uric acid, 69-93-2; xanthine, 69-89-6; carrier protein, 80700-39-6; Allantoin; Fungal Proteins; Membrane Transport Proteins; Uric Acid Funding details: Stavros Niarchos Foundation, SNF Funding text 1: We are grateful to George Lambrinidis for his help in making Figure 6. This work was supported by a Stavros S. Niarchos Foundation for Charity grant. Cited By :4 Export Date: 13 February 2020 CODEN: GENTA Correspondence Address: Diallinas, G.; Department of Biology, National and Kapodistrian University of Athens, Panepistimioupolis, Greece; email: diallina@biol.uoa.gr Chemicals/CAS: allantoin, 97-59-6; ammonia, 14798-03-9, 51847-23-5, 7664-41-7; uracil, 66-22-8; uric acid, 69-93-2; xanthine, 69-89-6; carrier protein, 80700-39-6; Allantoin; Fungal Proteins; Membrane Transport Proteins; Uric Acid Cited By :13 Export Date: 9 September 2021 CODEN: GENTA Correspondence Address: Diallinas, G.; Department of Biology, Panepistimioupolis, Greece; email: diallina@biol.uoa.gr Chemicals/CAS: allantoin, 97-59-6; ammonia, 14798-03-9, 51847-23-5, 7664-41-7; uracil, 66-22-8; uric acid, 69-93-2; xanthine, 69-89-6; carrier protein, 80700-39-6; Allantoin; Fungal Proteins; Membrane Transport Proteins; Uric Acid Funding details: Stavros Niarchos Foundation, SNF Funding text 1: We are grateful to George Lambrinidis for his help in making Figure 6. This work was supported by a Stavros S. Niarchos Foundation for Charity grant. LA - English DB - MTMT ER - TY - JOUR AU - Sorum, Ben AU - Törőcsik, Beáta AU - Csanády, László TI - Asymmetry of movements in CFTR's two ATP sites during pore opening serves their distinct functions JF - ELIFE J2 - ELIFE VL - 6 PY - 2017 PG - 17 SN - 2050-084X DO - 10.7554/eLife.29013 UR - https://m2.mtmt.hu/api/publication/3287727 ID - 3287727 AB - CFTR, the chloride channel mutated in cystic fibrosis (CF) patients, is opened by ATP binding to two cytosolic nucleotide binding domains (NBDs), but pore-domain mutations may also impair gating. ATP-bound NBDs dimerize occluding two nucleotides at interfacial binding sites; one site hydrolyzes ATP, the other is inactive. The pore opens upon tightening, and closes upon disengagement, of the catalytic site following ATP hydrolysis. Extent, timing, and role of non-catalytic-site movements are unknown. Here we exploit equilibrium gating of a hydrolysis-deficient mutant and apply Phi value analysis to compare timing of opening-associated movements at multiple locations, from the cytoplasmic ATP sites to the extracellular surface. Marked asynchrony of motion in the two ATP sites reveals their distinct roles in channel gating. The results clarify the molecular mechanisms of functional cross-talk between canonical and degenerate ATP sites in asymmetric ABC proteins, and of the gating defects caused by two common CF mutations. LA - English DB - MTMT ER - TY - JOUR AU - Timachi, MH AU - Hutter, CAJ AU - Hohl, M AU - Assafa, T AU - Böhm, S AU - Mittal, A AU - Seeger, MA AU - Bordignon, E TI - Exploring conformational equilibria of a heterodimeric ABC transporter JF - ELIFE J2 - ELIFE VL - 6 PY - 2017 SN - 2050-084X DO - 10.7554/eLife.20236 UR - https://m2.mtmt.hu/api/publication/26363440 ID - 26363440 N1 - Faculty of Chemistry and Biochemistry, Ruhr-Universität Bochum, Bochum, Germany Department of Physics, Freie Universität Berlin, Berlin, Germany Institute of Medical Microbiology, University of Zurich, Zurich, Switzerland Laboratory of Physical Chemistry, ETH Zurich, Zurich, Switzerland Department of Biochemistry, University of Zurich, Zurich, Switzerland Cited By :49 Export Date: 9 March 2022 Correspondence Address: Seeger, M.A.; Institute of Medical Microbiology, Switzerland; email: m.seeger@imm.uzh.ch Chemicals/CAS: adenosine triphosphatase, 37289-25-1, 9000-83-3; vanadic acid, 12260-63-8, 13981-20-9, 37353-31-4; adenosine triphosphate, 15237-44-2, 56-65-5, 987-65-5; Adenosine Triphosphate; ATP-Binding Cassette Transporters Funding details: Deutsche Forschungsgemeinschaft, DFG, BO3000/1?, EXC 1069 Funding details: Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung, SNF, PP00P3_144823 Funding details: Eidgenössische Technische Hochschule Zürich, ETH Funding details: Universität Zürich, UZH Funding text 1: EB would like to thank G Jeschke (ETH Zurich) for providing the Q-band resonator and R Schlesinger (FU Berlin) for providing access to the molecular biology lab. MAS thanks the Institute of Medical Microbiology and the University of Zurich for financial and administrative support. EB acknowledges support by the Deutsche Forschungsgemeinschaft (DFG) through grants BO3000/1?2 and INST 130/972?1 FUGG. This work is supported by the DFG Cluster of Excellence RESOLV (EXC 1069, to EB) funded by the Deutsche Forschungsgemeinschaft and by a SNF Professorship of the Swiss National Science Foundation (PP00P3_144823, to MAS). LA - English DB - MTMT ER -