Functional stability of CFTR depends on tight binding of ATP at its degenerate ATP‐binding site

Yeh, Han I.; Yu, Ying; Kuo, Pei Lun; Tsai, Chun Kuang; Huang, Hsin Tuan; Hwang, Tzyh Chang

doi:10.1113/jp281933

Cited by 11 publications

(6 citation statements)

References 71 publications

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“…7l ). Dissociation of ATP from both sites probably leads to the reversible rundown of CFTR currents that occurs after prolonged exposure to nucleotide-free solutions 29 .…”

Section: Dimerization Persists Through Cycles Of Hydrolysismentioning

confidence: 99%

CFTR function, pathology and pharmacology at single-molecule resolution

Levring

Terry

Kilic

et al. 2023

Nature

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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.

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“…7l ). Dissociation of ATP from both sites probably leads to the reversible rundown of CFTR currents that occurs after prolonged exposure to nucleotide-free solutions 29 .…”

Section: Dimerization Persists Through Cycles Of Hydrolysismentioning

confidence: 99%

CFTR function, pathology and pharmacology at single-molecule resolution

Levring

Terry

Kilic

et al. 2023

Nature

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“…The twelve transmembrane spans (TM) from the MSDs combine to form a pore allowing anions like chloride, bicarbonate but also iodide, to cross the PM. Only ATP binding site 2 has retained its ability to hydrolyze ATP, while site 1 was proposed to modulate the stability of CFTR function [ 28 ]. Dissociation of ADP in the NBD dimer after ATP hydrolysis at site 2 closes the channel (reviewed in [ 29 ]).…”

Section: One Size Does Not Fit Allmentioning

confidence: 99%

One Size Does Not Fit All: The Past, Present and Future of Cystic Fibrosis Causal Therapies

Ensinck

Carlon

2022

Cells

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Cystic fibrosis (CF) is the most common monogenic disorder, caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. Over the last 30 years, tremendous progress has been made in understanding the molecular basis of CF and the development of treatments that target the underlying defects in CF. Currently, a highly effective CFTR modulator treatment (Kalydeco™/Trikafta™) is available for 90% of people with CF. In this review, we will give an extensive overview of past and ongoing efforts in the development of therapies targeting the molecular defects in CF. We will discuss strategies targeting the CFTR protein (i.e., CFTR modulators such as correctors and potentiators), its cellular environment (i.e., proteostasis modulation, stabilization at the plasma membrane), the CFTR mRNA (i.e., amplifiers, nonsense mediated mRNA decay suppressors, translational readthrough inducing drugs) or the CFTR gene (gene therapies). Finally, we will focus on how these efforts can be applied to the 15% of people with CF for whom no causal therapy is available yet.

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“…Since these Lys and Gly are located at helix ends and not in loops, noisy fluctuations in the distance between their Cα atoms were prevented. These positions were identified by aligning the query NBDs to a reference NBD (Pgp NBD1 from 6c0v) using the TM-align algorithm and not by simple sequence search, since there are many ABC proteins with degenerate, non-conserved ATP-binding sites (e.g., CFTR/ABCC7 possesses a degenerate signature motif [ 45 , 46 ]). The distribution of the distances between Walker A and signature calculated for all structures ( Figure 3 ) indicated a cutoff value of 14 Å for distinguishing bottom-closed and bottom-open conformations.…”

Section: Resultsmentioning

confidence: 99%

Comprehensive Collection and Prediction of ABC Transmembrane Protein Structures in the AI Era of Structural Biology

Tordai

Suhajda

Sillitoe

et al. 2022

IJMS

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The number of unique transmembrane (TM) protein structures doubled in the last four years, which can be attributed to the revolution of cryo-electron microscopy. In addition, AlphaFold2 (AF2) also provided a large number of predicted structures with high quality. However, if a specific protein family is the subject of a study, collecting the structures of the family members is highly challenging in spite of existing general and protein domain-specific databases. Here, we demonstrate this and assess the applicability and usability of automatic collection and presentation of protein structures via the ABC protein superfamily. Our pipeline identifies and classifies transmembrane ABC protein structures using the PFAM search and also aims to determine their conformational states based on special geometric measures, conftors. Since the AlphaFold database contains structure predictions only for single polypeptide chains, we performed AF2-Multimer predictions for human ABC half transporters functioning as dimers. Our AF2 predictions warn of possibly ambiguous interpretation of some biochemical data regarding interaction partners and call for further experiments and experimental structure determination. We made our predicted ABC protein structures available through a web application, and we joined the 3D-Beacons Network to reach the broader scientific community through platforms such as PDBe-KB.

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Functional stability of CFTR depends on tight binding of ATP at its degenerate ATP‐binding site

Abstract: support-information-section).

Cited by 11 publications

References 71 publications

CFTR function, pathology and pharmacology at single-molecule resolution

CFTR function, pathology and pharmacology at single-molecule resolution

One Size Does Not Fit All: The Past, Present and Future of Cystic Fibrosis Causal Therapies

Comprehensive Collection and Prediction of ABC Transmembrane Protein Structures in the AI Era of Structural Biology

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