The cystic fibrosis mutation G1349D within the signature motif LSHGH of NBD2 abolishes the activation of CFTR chloride channels by genistein

Melin, Patricia; Thoreau, Vincent; Norez, Caroline; Bilan, Frédéric; Kitzis, Alain; Becq, Frédéric

doi:10.1016/j.bcp.2004.02.022

Cited by 36 publications

(46 citation statements)

References 31 publications

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“…Non-transfected CHO-K1, stably transfected wt-and delF508-CHO cells and Cos-7 cells transiently transfected with GFP-delF508-CFTR cDNA or GFP alone (denoted Cos-7 mock) were used as described [20,21].…”

Section: Cellsmentioning

confidence: 99%

Rescue of functional delF508‐CFTR channels in cystic fibrosis epithelial cells by the α‐glucosidase inhibitor miglustat

et al. 2006

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In the disease cystic fibrosis (CF), the most common mutation delF508 results in endoplasmic reticulum retention of misfolded CF gene proteins (CFTR). We show that the a-1,2-glucosidase inhibitor miglustat (N-butyldeoxynojirimycin, NB-DNJ) prevents delF508-CFTR/calnexin interaction and restores cAMP-activated chloride current in epithelial CF cells. Moreover, miglustat rescues a mature and functional delF508-CFTR in the intestinal crypts of ileal mucosa from delF508 mice. Since miglustat is an orally active orphan drug (Zavesca Ò ) prescribed for the treatment of Gaucher disease, our findings provide the basis for future clinical evaluation of miglustat in CF patients.

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Section: Cellsmentioning

confidence: 99%

Rescue of functional delF508‐CFTR channels in cystic fibrosis epithelial cells by the α‐glucosidase inhibitor miglustat

et al. 2006

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“…1) (11, 14). Substitution of this residue in the signature motif of NBD1 in ABCC7 (cystic fibrosis transmembrane conductance regulator) (Gly 551 ) causes severe cystic fibrosis (35). We therefore substituted aspartate for glycine in the signature motif of G5 and G8 (G5-G197D and G8-G216D) to investigate the possible role of this motif in protein function.…”

Section: -Azido[␣-mentioning

confidence: 99%

Functional Asymmetry of Nucleotide-binding Domains in ABCG5 and ABCG8

Zhang

Graf

Gerard

et al. 2006

Journal of Biological Chemistry

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The ATP-binding cassette half-transporters ABCG5 (G5) and ABCG8 (G8) promote secretion of neutral sterols into bile, a major pathway for elimination of sterols. Mutations in either ABCG5 or ABCG8 cause sitosterolemia, a recessive disorder characterized by impaired biliary and intestinal sterol secretion, sterol accumulation, and premature atherosclerosis. The mechanism by which the G5G8 heterodimer couples ATP hydrolysis to sterol transport is not known. Here we examined the roles of the Walker A, Walker B, and signature motifs in the nucleotide-binding domains (NBD) of G5 and G8 using recombinant adenoviruses to reconstitute biliary sterol transport in G5G8-deficient mice. Mutant forms of each halftransporter were co-expressed with their wild-type partners. Mutations at crucial residues in the Walker A and Walker B domains of G5 prevented biliary sterol secretion, whereas mutations of the corresponding residues in G8 did not. The opposite result was obtained when mutations were introduced into the signature motif; mutations in the signature domain of G8 prevented sterol transport, but substitution of the corresponding residues in G5 did not. Taken together, these findings indicate that the NBDs of G5 and G8 are not functionally equivalent. The integrity of the canonical NBD formed by the Walker A and Walker B motifs of G5 and the signature motif of G8 is essential for G5G8-mediated sterol transport. In contrast, mutations in key residues of the NBD formed by the Walker A and B motifs of G8 and the signature sequence of G5 did not affect sterol secretion. ABC2 transporters comprise a large family of polytopic membrane proteins that use the energy of ATP hydrolysis to translocate a wide variety of substrates across biological membranes (1). Most ABC transporters share a common architecture that includes two NBDs arranged in series with two membrane-spanning domains. The G family of ABC transporters includes five half-transporters that each contain a single hydrophilic NBD at the NH 2 terminus and a membrane-spanning domain consisting of six transmembrane ␣-helices at the COOH terminus. Some of the G family members, such as ABCG2, function as homodimers, whereas other family members, such as ABCG5 (G5) and ABCG8 (G8), function as heterodimers (2).G5 and G8 are expressed almost exclusively in hepatocytes and enterocytes, where they promote excretion of cholesterol and plant sterols into bile and into the gut lumen, respectively (3). Expression of the two proteins is coordinately regulated at the transcriptional level (3); ABCG5 and ABCG8 are juxtaposed on chromosome 2 and up-regulated by the nuclear hormone receptor liver X receptor (4), which promotes the expression of numerous other proteins involved in the centripetal movement of sterols from peripheral tissues to the liver (5). Expression of both G5 and G8 is required for either protein to be transported out of the endoplasmic reticulum (2, 6), and mutations in either ABCG5 or ABCG8 cause sitosterolemia, a rare autosomal recessive disorder characterized by ster...

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“…Despite the fact that numerous and diverse chemical scaffolds are now available, no pharmacophore has been identified, and surprisingly, very few drug binding sites have been mapped on the CFTR protein. However, studies with the CFTR activator isoflavonoid genistein highlighted the contribution of glycine residues Gly 551 and Gly 1349 of NBD1 and NBD2 domains, respectively, to the pharmacological modulation of CFTR by genistein (4,9). Mechanism of inhibition of CFTR activity by glibenclamide or CFTR inh -172 are associated with a positive charge of amino acids located on the third intracellular loop (Lys 918 ) and pore-forming transmembrane domains (Arg 347 ) respectively (4,10).…”

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confidence: 99%

C Terminus of Nucleotide Binding Domain 1 Contains Critical Features for Cystic Fibrosis Transmembrane Conductance Regulator Trafficking and Activation

Billet

Melin

Jollivet

et al. 2010

Journal of Biological Chemistry

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The cystic fibrosis transmembrane conductance regulator (CFTR) is a Cl؊ channel physiologically important in fluidtransporting epithelia and pathologically relevant in several human diseases. Here, we show that mutations in the C terminus of the first nucleotide binding domain comprising the latest ␤ strands (␤ c 5 and ␤ c 6) influence the trafficking, channel activity, and pharmacology of CFTR. We mutated CFTR amino acids located in the ␤ c 5-␤ c 6 hairpin, within the ␤ c 5 strand (H620Q), within the ␤-turn linking the two ␤ strands (E621G, G622D), as well as within (S623A, S624A) and at the extremity (G628R) of the ␤ c 6 strand. Functional analysis reveals that the current density was largely reduced for G622D and G628R channels compared with wt CFTR, similar for E621G and S624A, but increased for H620Q and S623A. For G622D and G628R, the abnormal activity is likely due to a defective maturation process, as assessed by the augmented activity and mature C-band observed in the presence of the trafficking corrector miglustat. In addition, in presence of the CFTR activator benzo[c]quinolizinium, the CFTR current density compared with that of wt CFTR was abolished for G622D and G628R channels, but similar for H620Q, S623A, and S624A or slightly increased for E621G. Finally, G622D and G628R were activated by the CFTR agonists genistein, RP-107, and isobutylmethylxanthine. Our results identify the C terminus of the CFTR first nucleotide binding domain as an important molecular site for the trafficking of CFTR protein, for the control of CFTR channel gating, and for the pharmacological effect of a dual activity agent.Mapping ligand binding sites within ionic channel structures is important to understand the mechanism of action of channel modulators (i.e. activators and inhibitors), to identify regions required for selective interaction with modulators, and to design more selective and more potent agents. Such mapping, called pharmaco-topology, has been performed for the sulfonylurea receptor (SUR) regulatory subunits of K ATP channels (reviewed in Ref.

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The cystic fibrosis mutation G1349D within the signature motif LSHGH of NBD2 abolishes the activation of CFTR chloride channels by genistein

Cited by 36 publications

References 31 publications

Rescue of functional delF508‐CFTR channels in cystic fibrosis epithelial cells by the α‐glucosidase inhibitor miglustat

Rescue of functional delF508‐CFTR channels in cystic fibrosis epithelial cells by the α‐glucosidase inhibitor miglustat

Functional Asymmetry of Nucleotide-binding Domains in ABCG5 and ABCG8

C Terminus of Nucleotide Binding Domain 1 Contains Critical Features for Cystic Fibrosis Transmembrane Conductance Regulator Trafficking and Activation

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