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EMBO J
2006 Oct 18;2520:4728-39. doi: 10.1038/sj.emboj.7601373.
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In vivo phosphorylation of CFTR promotes formation of a nucleotide-binding domain heterodimer.
Mense M
,
Vergani P
,
White DM
,
Altberg G
,
Nairn AC
,
Gadsby DC
.
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The human ATP-binding cassette (ABC) protein CFTR (cystic fibrosis transmembrane conductance regulator) is a chloride channel, whose dysfunction causes cystic fibrosis. To gain structural insight into the dynamic interaction between CFTR's nucleotide-binding domains (NBDs) proposed to underlie channel gating, we introduced target cysteines into the NBDs, expressed the channels in Xenopus oocytes, and used in vivo sulfhydryl-specific crosslinking to directly examine the cysteines' proximity. We tested five cysteine pairs, each comprising one introduced cysteine in the NH(2)-terminal NBD1 and another in the COOH-terminal NBD2. Identification of crosslinked product was facilitated by co-expression of NH(2)-terminal and COOH-terminal CFTR half channels each containing one NBD. The COOH-terminal half channel lacked all native cysteines. None of CFTR's 18 native cysteines was found essential for wild type-like, phosphorylation- and ATP-dependent, channel gating. The observed crosslinks demonstrate that NBD1 and NBD2 interact in a head-to-tail configuration analogous to that in homodimeric crystal structures of nucleotide-bound prokaryotic NBDs. CFTR phosphorylation by PKA strongly promoted both crosslinking and opening of the split channels, firmly linking head-to-tail NBD1-NBD2 association to channel opening.
Aleksandrov,
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.
2002, Pubmed
Aleksandrov,
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.
2002,
Pubmed
Basso,
Prolonged nonhydrolytic interaction of nucleotide with CFTR's NH2-terminal nucleotide binding domain and its role in channel gating.
2003,
Pubmed
,
Xenbase
Careaga,
Thermal motions of surface alpha-helices in the D-galactose chemosensory receptor. Detection by disulfide trapping.
1992,
Pubmed
Chan,
Severed molecules functionally define the boundaries of the cystic fibrosis transmembrane conductance regulator's NH(2)-terminal nucleotide binding domain.
2000,
Pubmed
,
Xenbase
Chang,
Protein kinase A (PKA) still activates CFTR chloride channel after mutagenesis of all 10 PKA consensus phosphorylation sites.
1993,
Pubmed
Chen,
A tweezers-like motion of the ATP-binding cassette dimer in an ABC transport cycle.
2003,
Pubmed
Csanády,
Severed channels probe regulation of gating of cystic fibrosis transmembrane conductance regulator by its cytoplasmic domains.
2000,
Pubmed
,
Xenbase
Csanády,
Preferential phosphorylation of R-domain Serine 768 dampens activation of CFTR channels by PKA.
2005,
Pubmed
,
Xenbase
Dassa,
The ABC of ABCS: a phylogenetic and functional classification of ABC systems in living organisms.
2001,
Pubmed
Dean,
The human ATP-binding cassette (ABC) transporter superfamily.
2001,
Pubmed
Denning,
Processing of mutant cystic fibrosis transmembrane conductance regulator is temperature-sensitive.
1992,
Pubmed
,
Xenbase
Fetsch,
Vanadate-catalyzed photocleavage of the signature motif of an ATP-binding cassette (ABC) transporter.
2002,
Pubmed
Gadsby,
Control of CFTR channel gating by phosphorylation and nucleotide hydrolysis.
1999,
Pubmed
Gregory,
Maturation and function of cystic fibrosis transmembrane conductance regulator variants bearing mutations in putative nucleotide-binding domains 1 and 2.
1991,
Pubmed
Higgins,
The ATP switch model for ABC transporters.
2004,
Pubmed
Hopfner,
Structural biology of Rad50 ATPase: ATP-driven conformational control in DNA double-strand break repair and the ABC-ATPase superfamily.
2000,
Pubmed
Hung,
Crystal structure of the ATP-binding subunit of an ABC transporter.
1998,
Pubmed
Karpowich,
Crystal structures of the MJ1267 ATP binding cassette reveal an induced-fit effect at the ATPase active site of an ABC transporter.
2001,
Pubmed
Kobashi,
Catalytic oxidation of sulfhydryl groups by o-phenanthroline copper complex.
1968,
Pubmed
Lewis,
Structure of nucleotide-binding domain 1 of the cystic fibrosis transmembrane conductance regulator.
2004,
Pubmed
Lewis,
Impact of the deltaF508 mutation in first nucleotide-binding domain of human cystic fibrosis transmembrane conductance regulator on domain folding and structure.
2005,
Pubmed
Liu,
Variable reactivity of an engineered cysteine at position 338 in cystic fibrosis transmembrane conductance regulator reflects different chemical states of the thiol.
2006,
Pubmed
,
Xenbase
Locher,
The E. coli BtuCD structure: a framework for ABC transporter architecture and mechanism.
2002,
Pubmed
Loo,
The "LSGGQ" motif in each nucleotide-binding domain of human P-glycoprotein is adjacent to the opposing walker A sequence.
2002,
Pubmed
Loo,
Membrane topology of a cysteine-less mutant of human P-glycoprotein.
1995,
Pubmed
Picciotto,
Phosphorylation of the cystic fibrosis transmembrane conductance regulator.
1992,
Pubmed
Pérez,
Reconstitution of expressed KCa channels from Xenopus oocytes to lipid bilayers.
1994,
Pubmed
,
Xenbase
Reyes,
Structure of the ABC transporter MsbA in complex with ADP.vanadate and lipopolysaccharide.
2005,
Pubmed
Riordan,
Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA.
1989,
Pubmed
Rosenberg,
Purification and crystallization of the cystic fibrosis transmembrane conductance regulator (CFTR).
2004,
Pubmed
Sali,
Comparative protein modelling by satisfaction of spatial restraints.
1993,
Pubmed
Samanta,
Disulfide cross-linking reveals a site of stable interaction between C-terminal regulatory domains of the two MalK subunits in the maltose transport complex.
2003,
Pubmed
Sheppard,
Structure and function of the CFTR chloride channel.
1999,
Pubmed
Smith,
ATP binding to the motor domain from an ABC transporter drives formation of a nucleotide sandwich dimer.
2002,
Pubmed
Stemmer,
Single-step assembly of a gene and entire plasmid from large numbers of oligodeoxyribonucleotides.
1995,
Pubmed
Szabó,
Nucleotide occlusion in the human cystic fibrosis transmembrane conductance regulator. Different patterns in the two nucleotide binding domains.
1999,
Pubmed
Thibodeau,
Side chain and backbone contributions of Phe508 to CFTR folding.
2005,
Pubmed
Verdon,
Crystal structures of the ATPase subunit of the glucose ABC transporter from Sulfolobus solfataricus: nucleotide-free and nucleotide-bound conformations.
2003,
Pubmed
Vergani,
On the mechanism of MgATP-dependent gating of CFTR Cl- channels.
2003,
Pubmed
,
Xenbase
Vergani,
CFTR channel opening by ATP-driven tight dimerization of its nucleotide-binding domains.
2005,
Pubmed
Wang,
Reversible silencing of CFTR chloride channels by glutathionylation.
2005,
Pubmed
Wu,
Helix proximity and ligand-induced conformational changes in the lactose permease of Escherichia coli determined by site-directed chemical crosslinking.
1997,
Pubmed
Yuan,
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.
2001,
Pubmed
Zaitseva,
H662 is the linchpin of ATP hydrolysis in the nucleotide-binding domain of the ABC transporter HlyB.
2005,
Pubmed
van Montfort,
Mapping of the dimer interface of the Escherichia coli mannitol permease by cysteine cross-linking.
2002,
Pubmed
