{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,9]],"date-time":"2026-01-09T02:20:58Z","timestamp":1767925258379,"version":"3.49.0"},"reference-count":42,"publisher":"Rockefeller University Press","issue":"4","content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2010,10,1]]},"abstract":"<jats:p>The chloride ion channel cystic fibrosis transmembrane conductance regulator (CFTR) displays a typical adenosine trisphosphate (ATP)-binding cassette (ABC) protein architecture comprising two transmembrane domains, two intracellular nucleotide-binding domains (NBDs), and a unique intracellular regulatory domain. Once phosphorylated in the regulatory domain, CFTR channels can open and close when supplied with cytosolic ATP. Despite the general agreement that formation of a head-to-tail NBD dimer drives the opening of the chloride ion pore, little is known about how ATP binding to individual NBDs promotes subsequent formation of this stable dimer. Structural studies on isolated NBDs suggest that ATP binding induces an intra-domain conformational change termed \u201cinduced fit,\u201d which is required for subsequent dimerization. We investigated the allosteric interaction between three residues within NBD2 of CFTR, F1296, N1303, and R1358, because statistical coupling analysis suggests coevolution of these positions, and because in crystal structures of ABC domains, interactions between these positions appear to be modulated by ATP binding. We expressed wild-type as well as F1296S, N1303Q, and R1358A mutant CFTR in Xenopus oocytes and studied these channels using macroscopic inside-out patch recordings. Thermodynamic mutant cycles were built on several kinetic parameters that characterize individual steps in the gating cycle, such as apparent affinities for ATP, open probabilities in the absence of ATP, open probabilities in saturating ATP in a mutant background (K1250R), which precludes ATP hydrolysis, as well as the rates of nonhydrolytic closure. Our results suggest state-dependent changes in coupling between two of the three positions (1296 and 1303) and are consistent with a model that assumes a toggle switch\u2013like interaction pattern during the intra-NBD2 induced fit in response to ATP binding. Stabilizing interactions of F1296 and N1303 present before ATP binding are replaced by a single F1296-N1303 contact in ATP-bound states, with similar interaction partner toggling occurring during the much rarer ATP-independent spontaneous openings.<\/jats:p>","DOI":"10.1085\/jgp.201010434","type":"journal-article","created":{"date-parts":[[2010,9,27]],"date-time":"2010-09-27T17:50:02Z","timestamp":1285609802000},"page":"407-423","update-policy":"https:\/\/doi.org\/10.1085\/jgp.crossmarkpolicy","source":"Crossref","is-referenced-by-count":23,"title":["Involvement of F1296 and N1303 of CFTR in induced-fit conformational change in response to ATP binding at NBD2"],"prefix":"10.1085","volume":"136","author":[{"given":"Andras","family":"Szollosi","sequence":"first","affiliation":[{"name":"Department of Medical Biochemistry, Semmelweis University, Budapest H-1094, Hungary 1"}]},{"given":"Paola","family":"Vergani","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Physiology and Pharmacology, University College London, London WC1E 7HY, England, UK 2"}]},{"given":"L\u00e1szl\u00f3","family":"Csan\u00e1dy","sequence":"additional","affiliation":[{"name":"Department of Medical Biochemistry, Semmelweis University, Budapest H-1094, Hungary 1"}]}],"member":"291","published-online":{"date-parts":[[2010,9,27]]},"reference":[{"key":"2023072521312291200_bib1","doi-asserted-by":"publisher","first-page":"15419","DOI":"10.1074\/jbc.M111713200","article-title":"The first nucleotide binding domain of cystic fibrosis transmembrane conductance regulator is a site of stable nucleotide interaction, whereas the second is a site of rapid turnover","volume":"277","author":"Aleksandrov","year":"2002","journal-title":"J. Biol. Chem."},{"key":"2023072521312291200_bib2","doi-asserted-by":"publisher","first-page":"333","DOI":"10.1085\/jgp.200308798","article-title":"Prolonged nonhydrolytic interaction of nucleotide with CFTR\u2019s NH2-terminal nucleotide binding domain and its role in channel gating","volume":"122","author":"Basso","year":"2003","journal-title":"J. Gen. Physiol."},{"key":"2023072521312291200_bib3","doi-asserted-by":"crossref","first-page":"2125","DOI":"10.1074\/jbc.M109539200","article-title":"Mutations that change the position of the putative gamma-phosphate linker in the nucleotide binding domains of CFTR alter channel gating","volume":"277","author":"Berger","year":"2002","journal-title":"J. Biol. Chem."},{"key":"2023072521312291200_bib4","doi-asserted-by":"publisher","first-page":"377","DOI":"10.1085\/jgp.200409228","article-title":"CFTR gating II: effects of nucleotide binding on the stability of open states","volume":"125","author":"Bompadre","year":"2005","journal-title":"J. Gen. Physiol."},{"key":"2023072521312291200_bib5","doi-asserted-by":"publisher","first-page":"605","DOI":"10.1085\/jgp.200308921","article-title":"Voltage-dependent gating of the cystic fibrosis transmembrane conductance regulator Cl\u2212 channel","volume":"122","author":"Cai","year":"2003","journal-title":"J. Gen. Physiol."},{"key":"2023072521312291200_bib6","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1085\/jgp.116.2.163","article-title":"Severed molecules functionally define the boundaries of the cystic fibrosis transmembrane conductance regulator\u2019s NH2-terminal nucleotide binding domain","volume":"116","author":"Chan","year":"2000","journal-title":"J. Gen. Physiol."},{"key":"2023072521312291200_bib7","doi-asserted-by":"publisher","first-page":"651","DOI":"10.1016\/j.molcel.2003.08.004","article-title":"A tweezers-like motion of the ATP-binding cassette dimer in an ABC transport cycle","volume":"12","author":"Chen","year":"2003","journal-title":"Mol. Cell."},{"key":"2023072521312291200_bib8","first-page":"483","article-title":"Fitting and statistical analysis of single-channel records","author":"Colquhoun","year":"1995"},{"key":"2023072521312291200_bib9","doi-asserted-by":"publisher","first-page":"129","DOI":"10.1085\/jgp.200910268","article-title":"Application of rate-equilibrium free energy relationship analysis to nonequilibrium ion channel gating mechanisms","volume":"134","author":"Csan\u00e1dy","year":"2009","journal-title":"J. Gen. Physiol."},{"key":"2023072521312291200_bib10","doi-asserted-by":"publisher","first-page":"477","DOI":"10.1085\/jgp.116.3.477","article-title":"Severed channels probe regulation of gating of cystic fibrosis transmembrane conductance regulator by its cytoplasmic domains","volume":"116","author":"Csan\u00e1dy","year":"2000","journal-title":"J. Gen. Physiol."},{"key":"2023072521312291200_bib11","doi-asserted-by":"publisher","first-page":"523","DOI":"10.1085\/jgp.200609558","article-title":"Thermodynamics of CFTR channel gating: a spreading conformational change initiates an irreversible gating cycle","volume":"128","author":"Csan\u00e1dy","year":"2006","journal-title":"J. Gen. Physiol."},{"key":"2023072521312291200_bib12","doi-asserted-by":"publisher","first-page":"1241","DOI":"10.1073\/pnas.0911061107","article-title":"Strict coupling between CFTR\u2019s catalytic cycle and gating of its Cl\u2212 ion pore revealed by distributions of open channel burst durations","volume":"107","author":"Csan\u00e1dy","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA."},{"key":"2023072521312291200_bib13","doi-asserted-by":"publisher","first-page":"477","DOI":"10.1038\/nature04712","article-title":"The ABC protein turned chloride channel whose failure causes cystic fibrosis","volume":"440","author":"Gadsby","year":"2006","journal-title":"Nature."},{"key":"2023072521312291200_bib14","doi-asserted-by":"publisher","first-page":"4964","DOI":"10.1093\/emboj\/20.17.4964","article-title":"Structure of the ABC ATPase domain of human TAP1, the transporter associated with antigen processing","volume":"20","author":"Gaudet","year":"2001","journal-title":"EMBO J."},{"key":"2023072521312291200_bib15","doi-asserted-by":"publisher","first-page":"1711","DOI":"10.1172\/JCI9622","article-title":"Inhibition of cystic fibrosis transmembrane conductance regulator by novel interaction with the metabolic sensor AMP-activated protein kinase","volume":"105","author":"Hallows","year":"2000","journal-title":"J. Clin. Invest."},{"key":"2023072521312291200_bib16","doi-asserted-by":"publisher","first-page":"412","DOI":"10.1016\/j.sbi.2007.07.003","article-title":"Structure and mechanism of ABC transporter proteins","volume":"17","author":"Hollenstein","year":"2007","journal-title":"Curr. Opin. Struct. Biol."},{"key":"2023072521312291200_bib17","doi-asserted-by":"publisher","first-page":"789","DOI":"10.1016\/S0092-8674(00)80890-9","article-title":"Structural biology of Rad50 ATPase: ATP-driven conformational control in DNA double-strand break repair and the ABC-ATPase superfamily","volume":"101","author":"Hopfner","year":"2000","journal-title":"Cell."},{"key":"2023072521312291200_bib18","doi-asserted-by":"publisher","first-page":"703","DOI":"10.1038\/25393","article-title":"Crystal structure of the ATP-binding subunit of an ABC transporter","volume":"396","author":"Hung","year":"1998","journal-title":"Nature."},{"key":"2023072521312291200_bib19","doi-asserted-by":"publisher","first-page":"571","DOI":"10.1016\/S0969-2126(01)00617-7","article-title":"Crystal structures of the MJ1267 ATP binding cassette reveal an induced-fit effect at the ATPase active site of an ABC transporter","volume":"9","author":"Karpowich","year":"2001","journal-title":"Structure."},{"key":"2023072521312291200_bib20","doi-asserted-by":"publisher","first-page":"5645","DOI":"10.1074\/jbc.M806780200","article-title":"Mechanistic insight into control of CFTR by AMPK","volume":"284","author":"Kongsuphol","year":"2009","journal-title":"J. Biol. Chem."},{"key":"2023072521312291200_bib21","doi-asserted-by":"publisher","first-page":"34711","DOI":"10.1074\/jbc.274.49.34711","article-title":"Large scale purification of detergent-soluble P-glycoprotein from Pichia pastoris cells and characterization of nucleotide binding properties of wild-type, Walker A, and Walker B mutant proteins","volume":"274","author":"Lerner-Marmarosh","year":"1999","journal-title":"J. Biol. Chem."},{"key":"2023072521312291200_bib22","doi-asserted-by":"publisher","first-page":"295","DOI":"10.1126\/science.286.5438.295","article-title":"Evolutionarily conserved pathways of energetic connectivity in protein families","volume":"286","author":"Lockless","year":"1999","journal-title":"Science."},{"key":"2023072521312291200_bib23","doi-asserted-by":"publisher","first-page":"2594","DOI":"10.1007\/s00018-008-8249-1","article-title":"Atomic model of human cystic fibrosis transmembrane conductance regulator: membrane-spanning domains and coupling interfaces","volume":"65","author":"Mornon","year":"2008","journal-title":"Cell. Mol. Life Sci."},{"key":"2023072521312291200_bib24","doi-asserted-by":"publisher","first-page":"1944","DOI":"10.1124\/mol.104.007708","article-title":"Functional interactions between nucleotide binding domains and leukotriene C4 binding sites of multidrug resistance protein 1 (ABCC1)","volume":"67","author":"Payen","year":"2005","journal-title":"Mol. Pharmacol."},{"key":"2023072521312291200_bib25","doi-asserted-by":"crossref","first-page":"12742","DOI":"10.1016\/S0021-9258(18)42339-3","article-title":"Phosphorylation of the cystic fibrosis transmembrane conductance regulator","volume":"267","author":"Picciotto","year":"1992","journal-title":"J. Biol. Chem."},{"key":"2023072521312291200_bib26","doi-asserted-by":"publisher","first-page":"15056","DOI":"10.1021\/bi061230e","article-title":"Thermodynamics of the ATPase cycle of GlcV, the nucleotide-binding domain of the glucose ABC transporter of sulfolobus solfataricus","volume":"45","author":"Pretz","year":"2006","journal-title":"Biochemistry."},{"key":"2023072521312291200_bib27","doi-asserted-by":"publisher","first-page":"51","DOI":"10.1016\/j.molcel.2006.07.034","article-title":"Distinct structural and functional properties of the ATPase sites in an asymmetric ABC transporter","volume":"24","author":"Procko","year":"2006","journal-title":"Mol. Cell."},{"key":"2023072521312291200_bib28","doi-asserted-by":"publisher","first-page":"1463","DOI":"10.1021\/bi982243y","article-title":"Walker mutations reveal loose relationship between catalytic and channel-gating activities of purified CFTR (cystic fibrosis transmembrane conductance regulator)","volume":"38","author":"Ramjeesingh","year":"1999","journal-title":"Biochemistry."},{"key":"2023072521312291200_bib29","doi-asserted-by":"publisher","first-page":"1066","DOI":"10.1126\/science.2475911","article-title":"Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA","volume":"245","author":"Riordan","year":"1989","journal-title":"Science."},{"key":"2023072521312291200_bib30","first-page":"700","article-title":"Single Channel Recording","author":"Sakmann","year":"1995","edition":"Second Edition"},{"key":"2023072521312291200_bib31","doi-asserted-by":"publisher","first-page":"139","DOI":"10.1016\/S1097-2765(02)00576-2","article-title":"ATP binding to the motor domain from an ABC transporter drives formation of a nucleotide sandwich dimer","volume":"10","author":"Smith","year":"2002","journal-title":"Mol. Cell."},{"key":"2023072521312291200_bib32","doi-asserted-by":"publisher","first-page":"628","DOI":"10.1038\/352628a0","article-title":"Phosphorylation-regulated Cl\u2212 channel in CHO cells stably expressing the cystic fibrosis gene","volume":"352","author":"Tabcharani","year":"1991","journal-title":"Nature."},{"key":"2023072521312291200_bib33","doi-asserted-by":"publisher","first-page":"405","DOI":"10.1085\/jgp.200810186","article-title":"State-dependent modulation of CFTR gating by pyrophosphate","volume":"133","author":"Tsai","year":"2009","journal-title":"J. Gen. Physiol."},{"key":"2023072521312291200_bib34","doi-asserted-by":"publisher","first-page":"11921","DOI":"10.1021\/bi001220s","article-title":"Investigation of the role of glutamine-471 and glutamine-1114 in the two catalytic sites of P-glycoprotein","volume":"39","author":"Urbatsch","year":"2000","journal-title":"Biochemistry."},{"key":"2023072521312291200_bib35","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1085\/jgp.104.1.123","article-title":"ATP alters current fluctuations of cystic fibrosis transmembrane conductance regulator: evidence for a three-state activation mechanism","volume":"104","author":"Venglarik","year":"1994","journal-title":"J. Gen. Physiol."},{"key":"2023072521312291200_bib36","doi-asserted-by":"publisher","first-page":"17","DOI":"10.1085\/jgp.20028673","article-title":"On the mechanism of MgATP-dependent gating of CFTR Cl\u2212 channels","volume":"121","author":"Vergani","year":"2003","journal-title":"J. Gen. Physiol."},{"key":"2023072521312291200_bib37","doi-asserted-by":"publisher","first-page":"876","DOI":"10.1038\/nature03313","article-title":"CFTR channel opening by ATP-driven tight dimerization of its nucleotide-binding domains","volume":"433","author":"Vergani","year":"2005","journal-title":"Nature."},{"key":"2023072521312291200_bib38","doi-asserted-by":"crossref","first-page":"945","DOI":"10.1002\/j.1460-2075.1982.tb01276.x","article-title":"Distantly related sequences in the alpha- and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold","volume":"1","author":"Walker","year":"1982","journal-title":"EMBO J."},{"key":"2023072521312291200_bib39","doi-asserted-by":"publisher","first-page":"55","DOI":"10.1085\/jgp.114.1.55","article-title":"Dual effects of ADP and adenylylimidodiphosphate on CFTR channel kinetics show binding to two different nucleotide binding sites","volume":"114","author":"Weinreich","year":"1999","journal-title":"J. Gen. Physiol."},{"key":"2023072521312291200_bib40","doi-asserted-by":"publisher","first-page":"1398","DOI":"10.1016\/S0006-3495(94)80930-0","article-title":"Effect of ATP concentration on CFTR Cl\u2212 channels: a kinetic analysis of channel regulation","volume":"66","author":"Winter","year":"1994","journal-title":"Biophys. J."},{"key":"2023072521312291200_bib41","doi-asserted-by":"publisher","first-page":"32313","DOI":"10.1074\/jbc.M100758200","article-title":"The crystal structure of the MJ0796 ATP-binding cassette. Implications for the structural consequences of ATP hydrolysis in the active site of an ABC transporter","volume":"276","author":"Yuan","year":"2001","journal-title":"J. Biol. Chem."},{"key":"2023072521312291200_bib42","doi-asserted-by":"publisher","first-page":"541","DOI":"10.1085\/jgp.113.4.541","article-title":"Gating of cystic fibrosis transmembrane conductance regulator chloride channels by adenosine triphosphate hydrolysis. Quantitative analysis of a cyclic gating scheme","volume":"113","author":"Zeltwanger","year":"1999","journal-title":"J. Gen. Physiol."}],"container-title":["Journal of General Physiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/rupress.org\/jgp\/article-pdf\/136\/4\/407\/1735226\/jgp_201010434.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/rupress.org\/jgp\/article-pdf\/136\/4\/407\/1735226\/jgp_201010434.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,7,25]],"date-time":"2023-07-25T21:32:01Z","timestamp":1690320721000},"score":1,"resource":{"primary":{"URL":"https:\/\/rupress.org\/jgp\/article\/136\/4\/407\/42869\/Involvement-of-F1296-and-N1303-of-CFTR-in-induced"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2010,9,27]]},"references-count":42,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2010,10,1]]}},"URL":"https:\/\/doi.org\/10.1085\/jgp.201010434","relation":{},"ISSN":["1540-7748","0022-1295"],"issn-type":[{"value":"1540-7748","type":"electronic"},{"value":"0022-1295","type":"print"}],"subject":[],"published":{"date-parts":[[2010,9,27]]}}}