CFTR Directly Mediates Nucleotide-regulated Glutathione Flux

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 2003 May 3

PMID 12727866

Citations 75

Authors

Ilana Kogan

Mohabir Ramjeesingh

Canhui Li

Jackie F Kidd

Yanchun Wang

Elaine M Leslie

Susan P C Cole

Christine E Bear

Affiliations

Soon will be listed here.

Abstract

Studies have shown that expression of cystic fibrosis transmembrane conductance regulator (CFTR) is associated with enhanced glutathione (GSH) efflux from airway epithelial cells, implicating a role for CFTR in the control of oxidative stress in the airways. To define the mechanism underlying CFTR-associated GSH flux, we studied wild-type and mutant CFTR proteins expressed in Sf9 membranes, as well as purified and reconstituted CFTR. We show that CFTR-expressing membrane vesicles mediate nucleotide-activated GSH flux, which is disrupted in the R347D pore mutant, and in the Walker A K464A and K1250A mutants. Further, we reveal that purified CFTR protein alone directly mediates nucleotide-dependent GSH flux. Interestingly, although ATP supports GSH flux through CFTR, this activity is enhanced in the presence of the non-hydrolyzable ATP analog AMP-PNP. These findings corroborate previous suggestions that CFTR pore properties can vary with the nature of the nucleotide interaction. In conclusion, our data demonstrate that GSH flux is an intrinsic function of CFTR and prompt future examination of the role of this function in airway biology in health and disease.

Citing Articles

Low Plasma Choline, High Trimethylamine Oxide, and Altered Phosphatidylcholine Subspecies Are Prevalent in Cystic Fibrosis Patients with Pancreatic Insufficiency.

Bernhard W, Shunova A, Boriga J, Graepler-Mainka U, Hilberath J Nutrients. 2025; 17(5).

PMID: 40077735 PMC: 11901616. DOI: 10.3390/nu17050868.

TNFRSF1B Signaling Blockade Protects Airway Epithelial Cells from Oxidative Stress.

Checa J, Fiol P, Guevara M, Aran J Antioxidants (Basel). 2024; 13(3).

PMID: 38539900 PMC: 10968085. DOI: 10.3390/antiox13030368.

Structural basis for autoinhibition by the dephosphorylated regulatory domain of Ycf1.

Khandelwal N, Tomasiak T Nat Commun. 2024; 15(1):2389.

PMID: 38493146 PMC: 10944535. DOI: 10.1038/s41467-024-46722-w.

Transcription factor BACH1 in cancer: roles, mechanisms, and prospects for targeted therapy.

Hu D, Zhang Z, Luo X, Li S, Jiang J, Zhang J Biomark Res. 2024; 12(1):21.

PMID: 38321558 PMC: 10848553. DOI: 10.1186/s40364-024-00570-4.

Cadmium transport by mammalian ATP-binding cassette transporters.

Thevenod F, Lee W Biometals. 2024; 37(3):697-719.

PMID: 38319451 PMC: 11101381. DOI: 10.1007/s10534-024-00582-5.

References

Linsdell P, Hanrahan J . Adenosine triphosphate-dependent asymmetry of anion permeation in the cystic fibrosis transmembrane conductance regulator chloride channel. J Gen Physiol. 1998; 111(4):601-14. PMC: 2217125. DOI: 10.1085/jgp.111.4.601. View

Egan M, Flotte T, Afione S, Solow R, Zeitlin P, Carter B . Defective regulation of outwardly rectifying Cl- channels by protein kinase A corrected by insertion of CFTR. Nature. 1992; 358(6387):581-4. DOI: 10.1038/358581a0. View

Wright S, Boag A, Valdimarsson G, Hipfner D, Campling B, Cole S . Immunohistochemical detection of multidrug resistance protein in human lung cancer and normal lung. Clin Cancer Res. 1998; 4(9):2279-89. View

Tsumura T, Hazama A, Miyoshi T, Ueda S, Okada Y . Activation of cAMP-dependent C1- currents in guinea-pig paneth cells without relevant evidence for CFTR expression. J Physiol. 1998; 512 ( Pt 3):765-77. PMC: 2231250. DOI: 10.1111/j.1469-7793.1998.765bd.x. View

Marvao P, de Jesus Ferreira M, Bailly C, Paulais M, Bens M, Guinamard R . Cl- absorption across the thick ascending limb is not altered in cystic fibrosis mice. A role for a pseudo-CFTR Cl- channel. J Clin Invest. 1998; 102(11):1986-93. PMC: 509151. DOI: 10.1172/JCI4074. View

Ramjeesingh M, Garami E, Galley K, Li C, Wang Y, Bear C . Purification and reconstitution of epithelial chloride channel cystic fibrosis transmembrane conductance regulator. Methods Enzymol. 1999; 294:227-46. DOI: 10.1016/s0076-6879(99)94014-4. View

Lee M, Wigley W, Zeng W, Noel L, Marino C, Thomas P . Regulation of Cl-/ HCO3- exchange by cystic fibrosis transmembrane conductance regulator expressed in NIH 3T3 and HEK 293 cells. J Biol Chem. 1999; 274(6):3414-21. DOI: 10.1074/jbc.274.6.3414. View

Frizzell R . Ten years with CFTR. Physiol Rev. 1999; 79(1 Suppl):S1-2. DOI: 10.1152/physrev.1999.79.1.S1. View

Schwiebert E, Benos D, Egan M, Stutts M, Guggino W . CFTR is a conductance regulator as well as a chloride channel. Physiol Rev. 1999; 79(1 Suppl):S145-66. DOI: 10.1152/physrev.1999.79.1.S145. View

10.

Ramjeesingh M, Li C, Garami E, Huan L, Galley K, Wang Y . Walker mutations reveal loose relationship between catalytic and channel-gating activities of purified CFTR (cystic fibrosis transmembrane conductance regulator). Biochemistry. 1999; 38(5):1463-8. DOI: 10.1021/bi982243y. View

11.

Cotten J, Welsh M . Cystic fibrosis-associated mutations at arginine 347 alter the pore architecture of CFTR. Evidence for disruption of a salt bridge. J Biol Chem. 1999; 274(9):5429-35. DOI: 10.1074/jbc.274.9.5429. View

12.

Boucher R . Molecular insights into the physiology of the 'thin film' of airway surface liquid. J Physiol. 1999; 516 ( Pt 3):631-8. PMC: 2269304. DOI: 10.1111/j.1469-7793.1999.0631u.x. View

13.

Gao L, Kim K, Yankaskas J, Forman H . Abnormal glutathione transport in cystic fibrosis airway epithelia. Am J Physiol. 1999; 277(1):L113-8. DOI: 10.1152/ajplung.1999.277.1.L113. View

14.

Kelly F . Gluthathione: in defence of the lung. Food Chem Toxicol. 1999; 37(9-10):963-6. DOI: 10.1016/s0278-6915(99)00087-3. View

15.

Hwang T, Sheppard D . Molecular pharmacology of the CFTR Cl- channel. Trends Pharmacol Sci. 1999; 20(11):448-53. DOI: 10.1016/s0165-6147(99)01386-3. View

16.

Ikuma M, Welsh M . Regulation of CFTR Cl- channel gating by ATP binding and hydrolysis. Proc Natl Acad Sci U S A. 2000; 97(15):8675-80. PMC: 27007. DOI: 10.1073/pnas.140220597. View

17.

Loe D, Oleschuk C, Deeley R, Cole S . Structure-activity studies of verapamil analogs that modulate transport of leukotriene C(4) and reduced glutathione by multidrug resistance protein MRP1. Biochem Biophys Res Commun. 2000; 275(3):795-803. DOI: 10.1006/bbrc.2000.3384. View

18.

Rahman I, MacNee W . Oxidative stress and regulation of glutathione in lung inflammation. Eur Respir J. 2000; 16(3):534-54. DOI: 10.1034/j.1399-3003.2000.016003534.x. View

19.

Fuller C, Ji H, Tousson A, Elble R, Pauli B, Benos D . Ca(2+)-activated Cl(-) channels: a newly emerging anion transport family. Pflugers Arch. 2002; 443 Suppl 1:S107-10. DOI: 10.1007/s004240100655. View

20.

Reddy M, Quinton P . Selective activation of cystic fibrosis transmembrane conductance regulator Cl- and HCO3- conductances. JOP. 2002; 2(4 Suppl):212-8. View