Conformational Changes Relevant to Channel Activity and Folding within the first Nucleotide Binding Domain of the Cystic Fibrosis Transmembrane Conductance Regulator

Hudson, Rhea; Chong, P. Andrew; Protasevich, I.I.; Vernon, Robert; Noy, Efrat; Bihler, Hermann; An, Jian Li; Kalid, Ori; Sela-Culang, Inbal; Mense, Martin; Senderowitz, Hanoch; Brouillette, Christie G.; Forman-Kay, Julie D.

doi:10.1074/jbc.m112.371138

Cited by 47 publications

(75 citation statements)

References 63 publications

(77 reference statements)

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“…Hydrogen/deuterium (H/D) exchange experiments show that the amide protons in this a-helix readily exchange in solution, indicating that the helix is unstable . NMR-derived chemical shifts provide evidence that the preceding two helices (H8 and H9) also exchange with a coil conformation (Hudson et al 2012). Crystal structures show short helices at both the amino and carboxyl termini of the RI; however, electron density is absent for the central portion, implying that this region of the RI is disordered.…”

Section: Nbd1mentioning

confidence: 97%

Dynamics Intrinsic to Cystic Fibrosis Transmembrane Conductance Regulator Function and Stability

Chong

Kota

Dokholyan

et al. 2013

Cold Spring Harbor Perspectives in Medicine

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The cystic fibrosis transmembrane conductance regulator (CFTR) requires dynamic fluctuations between states in its gating cycle for proper channel function, including changes in the interactions between the nucleotide-binding domains (NBDs) and between the intracellular domain (ICD) coupling helices and NBDs. Such motions are also linked with fluctuating phosphorylation-dependent binding of CFTR's disordered regulatory (R) region to the NBDs and partners. Folding of CFTR is highly inefficient, with the marginally stable NBD1 sampling excited states or folding intermediates that are aggregation-prone. The severe CFcausing F508del mutation exacerbates the folding inefficiency of CFTR and leads to impaired channel regulation and function, partly as a result of perturbed NBD1 -ICD interactions and enhanced sampling of these NBD1 excited states. Increased knowledge of the dynamics within CFTR will expand our understanding of the regulated channel gating of the protein as well as of the F508del defects in folding and function.

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Section: Nbd1mentioning

confidence: 97%

Dynamics Intrinsic to Cystic Fibrosis Transmembrane Conductance Regulator Function and Stability

Chong

Kota

Dokholyan

et al. 2013

Cold Spring Harbor Perspectives in Medicine

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“…4) shows that R region binding causes moderate, but significant, chemical shift changes throughout NBD1 (Fig. 4B) using previously obtained assignments for NBD1 (27). These wide-spread chemical shifts are likely because of the shallow energy landscape of NBD1 that facilitates allosteric effects caused by the binding (27,28), and they are difficult to simply interpret in terms of direct contacts.…”

Section: R Region Binds Multiple Partners In a Phosphorylation-dependentmentioning

confidence: 99%

“…4B) using previously obtained assignments for NBD1 (27). These wide-spread chemical shifts are likely because of the shallow energy landscape of NBD1 that facilitates allosteric effects caused by the binding (27,28), and they are difficult to simply interpret in terms of direct contacts. Resonance broadening, however, is localized primarily to two distinct sites around residues 593 and 618, suggesting that these represent effects of direct interaction (Fig.…”

Section: R Region Binds Multiple Partners In a Phosphorylation-dependentmentioning

confidence: 99%

Regulatory R region of the CFTR chloride channel is a dynamic integrator of phospho-dependent intra- and intermolecular interactions

Bozóky

Krzeminski

Muhandiram

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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151

218

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Intrinsically disordered proteins play crucial roles in regulatory processes and often function as protein interaction hubs. Here, we present a detailed characterization of a full-length disordered hub protein region involved in multiple dynamic complexes. We performed NMR, CD, and fluorescence binding studies on the nonphosphorylated and highly PKA-phosphorylated human cystic fibrosis transmembrane conductance regulator (CFTR) regulatory region, a ∼200-residue disordered segment involved in phosphorylation-dependent regulation of channel trafficking and gating. Our data provide evidence for dynamic, phosphorylation-dependent, multisite interactions of various segments of the regulatory region for its intra-and intermolecular partners, including the CFTR nucleotide binding domains 1 and 2, a 42-residue peptide from the C terminus of CFTR, the SLC26A3 sulphate transporter and antisigma factor antagonist (STAS) domain, and 14-3-3β. Because of its large number of binding partners, multivalent binding of individually weak sites facilitates rapid exchange between free and bound states to allow the regulatory region to engage with different partners and generate a graded or rheostat-like response to phosphorylation. Our results enrich the understanding of how disordered binding segments interact with multiple targets. We present structural models consistent with our data that illustrate this dynamic aspect of phospho-regulation of CFTR by the disordered regulatory region.protein interaction network | fuzzy complex

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“…1A). Transmembrane helix predictions (50), sequence alignments (38,44), and homology models (44) of SUR2B based on structures of other ABC proteins (51-57) indicate that transmembrane helix 11 ends at Val-599, that residues Gln-600 -Leu-607 extend transmembrane helix 11 into the cytoplasm, and that NBD1 begins at Asp-665 (Fig. 1A).…”

Section: Identification Of Phosphorylation Sites In Sur2b Nbd1-mentioning

confidence: 99%

Phosphorylation-dependent Changes in Nucleotide Binding, Conformation, and Dynamics of the First Nucleotide Binding Domain (NBD1) of the Sulfonylurea Receptor 2B (SUR2B)

Araujo

Alvarez

López‐Alonso

et al. 2015

Journal of Biological Chemistry

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Conformational Changes Relevant to Channel Activity and Folding within the first Nucleotide Binding Domain of the Cystic Fibrosis Transmembrane Conductance Regulator

Cited by 47 publications

References 63 publications

Dynamics Intrinsic to Cystic Fibrosis Transmembrane Conductance Regulator Function and Stability

Dynamics Intrinsic to Cystic Fibrosis Transmembrane Conductance Regulator Function and Stability

Regulatory R region of the CFTR chloride channel is a dynamic integrator of phospho-dependent intra- and intermolecular interactions

Phosphorylation-dependent Changes in Nucleotide Binding, Conformation, and Dynamics of the First Nucleotide Binding Domain (NBD1) of the Sulfonylurea Receptor 2B (SUR2B)

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